<?xml version="1.0" encoding="UTF-8" standalone="no"?>
<!DOCTYPE article PUBLIC "-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" "journalpublishing.dtd">
<article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" article-type="review-article" dtd-version="2.3" xml:lang="EN">
<front>
<journal-meta>
<journal-id journal-id-type="publisher-id">Front. Cell. Infect. Microbiol.</journal-id>
<journal-title>Frontiers in Cellular and Infection Microbiology</journal-title>
<abbrev-journal-title abbrev-type="pubmed">Front. Cell. Infect. Microbiol.</abbrev-journal-title>
<issn pub-type="epub">2235-2988</issn>
<publisher>
<publisher-name>Frontiers Media S.A.</publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="doi">10.3389/fcimb.2022.1012533</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Cellular and Infection Microbiology</subject>
<subj-group>
<subject>Mini Review</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group>
<article-title>Modification of phosphoinositides by the <italic>Shigella</italic> effector IpgD during host cell infection</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author" corresp="yes">
<name>
<surname>Tran Van Nhieu</surname>
<given-names>Guy</given-names>
</name>
<xref ref-type="aff" rid="aff1">
<sup>1</sup>
</xref>
<xref ref-type="author-notes" rid="fn001">
<sup>*</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/1997567"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Latour-Lambert</surname>
<given-names>Patricia</given-names>
</name>
<xref ref-type="aff" rid="aff2">
<sup>2</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/1943488"/>
</contrib>
<contrib contrib-type="author" corresp="yes">
<name>
<surname>Enninga</surname>
<given-names>Jost</given-names>
</name>
<xref ref-type="aff" rid="aff2">
<sup>2</sup>
</xref>
<xref ref-type="author-notes" rid="fn001">
<sup>*</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/16243"/>
</contrib>
</contrib-group>
<aff id="aff1">
<sup>1</sup>
<institution>Institute for Integrative Biology of the Cell &#x2013; Centre National de la Recherche Scientifique (CNRS) UMR9198 - Institut National de la Sant&#xe9; et de la Recherche M&#xe9;dicale (Inserm) U1280, Team Calcium Signaling and Microbial Infections</institution>, <addr-line>Gif-sur-Yvette</addr-line>, <country>France</country>
</aff>
<aff id="aff2">
<sup>2</sup>
<institution>Institut Pasteur, Unit&#xe9; Dynamique des interactions h&#xf4;tes-pathog&#xe8;nes and Centre National de la Recherche Scientifique (CNRS) UMR3691, Universit&#xe9; de Paris Cit&#xe9;</institution>, <addr-line>Paris</addr-line>, <country>France</country>
</aff>
<author-notes>
<fn fn-type="edited-by">
<p>Edited by: Gerardo Daniel Fidelio, National University of Cordoba (CIQUIBIC), Argentina</p>
</fn>
<fn fn-type="edited-by">
<p>Reviewed by: Juan Jose Martinez, Louisiana State University, United States</p>
</fn>
<fn fn-type="corresp" id="fn001">
<p>*Correspondence: Guy Tran Van Nhieu, <email xlink:href="mailto:guy.tranvannhieul@i2bc.paris-saclay.fr">guy.tranvannhieul@i2bc.paris-saclay.fr</email>; Jost Enninga, <email xlink:href="mailto:jost.enninga@pasteur.fr">jost.enninga@pasteur.fr</email>
</p>
</fn>
<fn fn-type="other" id="fn002">
<p>This article was submitted to Molecular Bacterial Pathogenesis, a section of the journal Frontiers in Cellular and Infection Microbiology</p>
</fn>
</author-notes>
<pub-date pub-type="epub">
<day>27</day>
<month>10</month>
<year>2022</year>
</pub-date>
<pub-date pub-type="collection">
<year>2022</year>
</pub-date>
<volume>12</volume>
<elocation-id>1012533</elocation-id>
<history>
<date date-type="received">
<day>05</day>
<month>08</month>
<year>2022</year>
</date>
<date date-type="accepted">
<day>27</day>
<month>09</month>
<year>2022</year>
</date>
</history>
<permissions>
<copyright-statement>Copyright &#xa9; 2022 Tran Van Nhieu, Latour-Lambert and Enninga</copyright-statement>
<copyright-year>2022</copyright-year>
<copyright-holder>Tran Van Nhieu, Latour-Lambert and Enninga</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/">
<p>This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</p>
</license>
</permissions>
<abstract>
<p>
<italic>Shigella</italic>, the causative agent of bacillary dysentery, subvert cytoskeletal and trafficking processes to invade and replicate in epithelial cells using an arsenal of bacterial effectors translocated through a type III secretion system. Here, we review the various roles of the type III effector IpgD, initially characterized as phosphatidylinositol 4,5 bisphosphate (PI4,5P<sub>2</sub>) 4-phosphatase. By decreasing PI4,5P<sub>2</sub> levels, IpgD triggers the disassembly of cortical actin filaments required for bacterial invasion and cell migration. PI5P produced by IpgD further stimulates signaling pathways regulating cell survival, macropinosome formation, endosomal trafficking and dampening of immune responses. Recently, IpgD was also found to exhibit phosphotransferase activity leading to PI3,4P<sub>2</sub> synthesis adding a new flavor to this multipotent bacterial enzyme. The substrate of IpgD, PI4,5P<sub>2</sub> is also the main substrate hydrolyzed by endogenous phospholipases C to produce inositoltriphosphate (InsP<sub>3</sub>), a major Ca<sup>2+</sup> second messenger. Hence, beyond the repertoire of effects associated with the direct diversion of phoshoinositides, IpgD indirectly down-regulates InsP<sub>3</sub>-mediated Ca<sup>2+</sup> release by limiting InsP<sub>3</sub> production. Furthermore, IpgD controls the intracellular lifestyle of <italic>Shigella</italic> promoting Rab8/11 -dependent recruitment of the exocyst at macropinosomes to remove damaged vacuolar membrane remnants and promote bacterial cytosolic escape. IpgD thus emerges as a key bacterial effector for the remodeling of host cell membranes.</p>
</abstract>
<kwd-group>
<kwd>
<italic>Shigella</italic>
</kwd>
<kwd>IpgD</kwd>
<kwd>phosphatidyl-inositol polyphosphate</kwd>
<kwd>phosphatase</kwd>
<kwd>invasion</kwd>
<kwd>dissemination</kwd>
<kwd>inflammation</kwd>
<kwd>phosphotransferase</kwd>
</kwd-group>
<contract-sponsor id="cn001">European Research Council<named-content content-type="fundref-id">10.13039/501100000781</named-content>
</contract-sponsor>
<contract-sponsor id="cn002">Agence Nationale de la Recherche<named-content content-type="fundref-id">10.13039/501100001665</named-content>
</contract-sponsor>
<contract-sponsor id="cn003">Labex<named-content content-type="fundref-id">10.13039/501100004100</named-content>
</contract-sponsor>
<contract-sponsor id="cn004">Labex<named-content content-type="fundref-id">10.13039/501100004100</named-content>
</contract-sponsor>
<contract-sponsor id="cn005">Agence Nationale de la Recherche<named-content content-type="fundref-id">10.13039/501100001665</named-content>
</contract-sponsor>
<counts>
<fig-count count="2"/>
<table-count count="0"/>
<equation-count count="0"/>
<ref-count count="63"/>
<page-count count="9"/>
<word-count count="4150"/>
</counts>
</article-meta>
</front>
<body>
<sec id="s1" sec-type="intro">
<title>Introduction</title>
<p>Invasive pathogens subvert cytoskeletal processes to promote their internalization by normally non-phagocytic cells. Once inside a targeted host cell, the pathogens may reside inside a membrane-bound bacterial-containing vacuole (BCV) or escape from this compartment. Diversion of endocytic processes has been mostly described for pathogens living inside BCVs. Recent evidence indicates that freely replicating bacteria within the cell cytosol, such as <italic>Shigella</italic>, the causative agent of bacillary dysentery, also divert host trafficking to promote vacuolar escape. Because of their central role in cytoskeletal reorganization and endosomal trafficking, phosphoinositides are targeted by virulence factors from a variety of invasive bacteria (<xref ref-type="bibr" rid="B40">Payrastre et&#xa0;al., 2012</xref>; <xref ref-type="bibr" rid="B43">Pizarro-Cerd&#xe1; et&#xa0;al., 2015</xref>). Among these, type III secretion system (T3SS) effectors that are injected into the host cell cytosol are of particular interest. They alter general cellular signaling pathways, as well as exhibiting localized activities at bacterial entry sites (<xref ref-type="bibr" rid="B22">Jennings et&#xa0;al., 2017</xref>; <xref ref-type="bibr" rid="B36">Muthuramalingam et&#xa0;al., 2021</xref>).</p>
<p>The <italic>Shigella</italic> IpgD type III effector has been characterized in detail as a PIP<sub>2</sub> 4-phosphatase involved in key processes during host cell infection (<xref ref-type="bibr" rid="B40">Payrastre et&#xa0;al., 2012</xref>). IpgD-mediated effects have been linked to its PIP<sub>2</sub> hydrolytic activity, decreased inositol(1, 4, 5) triphosphate (InsP<sub>3</sub>) levels and increased production of PI5P (<xref ref-type="bibr" rid="B40">Payrastre et&#xa0;al., 2012</xref>). In a recent study, IpgD has been shown- similar to the <italic>Salmonella</italic> T3SS ortholog SopB to act as a phosphotransferase producing PI3,4P<sub>2</sub> questioning the precise enzymatic activity pertinent to host cell process subversion (<xref ref-type="bibr" rid="B58">Walpole et&#xa0;al., 2022</xref>). IpgD modulates local signals occurring at bacterial-host cell contact sites during invasion and during rupture of the phagocytic vacuole by regulating the activity of small GTPases and specific phosphoinositide levels associated with the BCV (<xref ref-type="bibr" rid="B30">L&#xf3;pez-Montero and Enninga, 2018</xref>; <xref ref-type="bibr" rid="B34">Morioka et&#xa0;al., 2018</xref>). It also regulates Ca<sup>2+</sup> signaling by depleting the second messenger InsP<sub>3</sub> from T3SS-targeted cells (<xref ref-type="bibr" rid="B53">Sun et&#xa0;al., 2017</xref>) Furthermore, IpgD is the main inducer of macropinosomes surrounding invading <italic>Shigella</italic> (<xref ref-type="bibr" rid="B63">Weiner et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B8">Chang et&#xa0;al., 2020</xref>). The kinetics of (i) invasion and (ii) vacuolar rupture are tightly linked, suggesting an interdependence between both processes (<xref ref-type="bibr" rid="B12">Enninga et&#xa0;al., 2005</xref>). Due to the overall complexity of the process, it remains poorly understood how IpgD mechanistically contributes to vacuolar rupture to promote <italic>Shigella</italic> intracellular replication. Additional T3SS effectors, such as IpgB1/2, IpaJ, VirB and IcsB, also modulate the dynamics of host cell membranes during bacterial entry (<xref ref-type="bibr" rid="B16">Handa et&#xa0;al., 2007</xref>; <xref ref-type="bibr" rid="B15">Hachani et&#xa0;al., 2008</xref>; <xref ref-type="bibr" rid="B17">Heindl et&#xa0;al., 2010</xref>; <xref ref-type="bibr" rid="B6">Burnaevskiy et&#xa0;al., 2013</xref>; <xref ref-type="bibr" rid="B27">K&#xfc;hn et&#xa0;al., 2020</xref>).</p>
<p>IpgD thus exhibits multiple effects on phosphoinositides as well as on Ca<sup>2+</sup> signaling. Here, we review the diverse processes targeted by IpgD with a particular emphasis on findings implicating IpgD in the diversion of macropinosome formation and endosomal trafficking leading to rupture of the bacterial containing vacuole.</p>
</sec>
<sec id="s2">
<title>Global IpgD-mediated effects on cell signaling and inflammation</title>
<p>Through its enzymatic activities, IpgD regulates many aspects of signaling involved in survival pathways and inflammatory processes. IpgD leads to the production of PI5P at <italic>Shigella</italic> invasion sites inducing the tyrosine kinase-dependent activation of class IA PI3K and activation of the Akt serine/threonine kinase regulating cell survival and proliferation (<xref ref-type="bibr" rid="B41">Pendaries et&#xa0;al., 2006</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). In light of the recently described IpgD phosphotransferase activity, it is possible that Akt activation also occurs directly <italic>via</italic> increased PI 3,4P<sub>2</sub> levels triggered by IpgD (<xref ref-type="bibr" rid="B58">Walpole et&#xa0;al., 2022</xref>). Contributing to cell survival, IpgD specifically affects the dynamics of membranes at the host cell surface including endosome and macropinosome formation. Through increased PI5P levels at the plasma membrane of infected cells and a yet to be described mechanism, IpgD stimulates endocytosis of the EGF receptor (EGFR) and the cell adhesion molecule ICAM-1 (<xref ref-type="bibr" rid="B2">Boal et&#xa0;al., 2016</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). While internalization is increased for these cell surface proteins, their outcome differs suggesting differential and specific cargo sorting for EGFR and ICAM-1 endosomes (<xref ref-type="bibr" rid="B46">Ramel et&#xa0;al., 2011</xref>; <xref ref-type="bibr" rid="B2">Boal et&#xa0;al., 2016</xref>). IpgD prevents the degradation of EGFR by blocking endosome maturation through the PI5P-dependent recruitment of the TOM1 adaptor protein at endosomal membranes (<xref ref-type="bibr" rid="B1">Boal et&#xa0;al., 2015</xref>). This results in the persistence of EGFR-early endosomes and persistent EGFR-signaling favoring the PI3K-Akt survival pathway (<xref ref-type="bibr" rid="B1">Boal et&#xa0;al., 2015</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). The precise mechanism involved in the PI5P/TOM1-dependent blockade of endosome maturation remains to be characterized and appears unrelated to its described function in endosomal sorting. In contrast, as expected from canonical endosomal maturation, the increased internalization of ICAM-1 leads to its targeting to lysosomes and degradation (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). As a global result, IpgD stimulates the removal of ICAM-1 from the epithelial cell surface, thereby reducing neutrophil adhesion and clearance of infected cells (<xref ref-type="bibr" rid="B2">Boal et&#xa0;al., 2016</xref>). Through PI5P, IpgD also prevents the recruitment of immune cells and pro-inflammatory signals by limiting the release of extracellular ATP (eATP) from infected cells (<xref ref-type="bibr" rid="B45">Puhar et&#xa0;al., 2013</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). Under basal conditions the eATP concentration is negligible, while being estimated in the order of 5-10 mM in the cytosol of host cells. Therefore, eATP is a well-characterized danger signal associated with cell injury, activation of pro-inflammatory cytokines and immune cell recruitment. Bacteria, such as <italic>Shigella</italic>, expressing a T3SS induce the release of eATP at the early stages of infection in the absence of detectable host cell lysis (<xref ref-type="bibr" rid="B45">Puhar et&#xa0;al., 2013</xref>). <italic>Shigella</italic>-induced eATP release results from an increase of cytosolic Ca<sup>2+</sup> levels in infected cells leading to the opening of connexin hemichannels (<xref ref-type="bibr" rid="B45">Puhar et&#xa0;al., 2013</xref>). PI5P produced by IpgD prevents connexin hemichannel opening and the release of eATP associated with <italic>Shigella</italic> invasion, thereby dampening associated immune responses (<xref ref-type="bibr" rid="B45">Puhar et&#xa0;al., 2013</xref>).</p>
<fig id="f1" position="float">
<label>Figure&#xa0;1</label>
<caption>
<p>Versatile roles of IpgD during epithelial cell infection. Through PI4,5P<sub>2</sub> (PIP<sub>2</sub>) hydrolysis, IpgD disconnect cortical actin from plasma membranes, favoring bacterial invasion and cell to cell spread. IpgD-mediated production of PI5P stimulates actin polymerization <italic>via</italic> Tiam-Rac activation, down-regulates the recruitment of immune cells by inducing ICAM-1 degradation, prevents the opening of connexin hemichannels, promotes persistent EGFR signaling by recruiting the TOM1 adaptor to endosome and preventing their maturation, and activates cell survival through the PI3K/Akt pathway. Possibly, these effects could also be linked to increased PI3,4P2 levels associated with the IpgD phosphotransferase activity. By decreasing InsP<sub>3</sub> levels, IpgD allows long lasting local Ca<sup>2+</sup> responses during invasion and prevents InsP<sub>3</sub>-dependent global Ca<sup>2+</sup> responses at later infection stages. Inhibition of Ca2+ signaling and calpain activity favors adhesion of infected cells. Through mechanisms the remain to be characterized, IpgD stimulates the formation of macropinosomes during bacterial invasion. IpgD also allows the recruitment of Rab11-positive endosomes and the exocyst, triggering bacterial unsheathing from the actin cocoon surrounding the BCV and vacuolar escape.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fcimb-12-1012533-g001.tif"/>
</fig>
<p>Finally, <italic>Shigella</italic> negatively regulates the recruitment of neutrophils at infection sites through direct bacterial contact with the immune cells (<xref ref-type="bibr" rid="B25">Konradt et&#xa0;al., 2011</xref>). While initially described during intestinal epithelial infection, it was also reported that injection of T3SS effectors occurred in neutrophils by transiently interacting bacteria in the absence of bacterial invasion (<xref ref-type="bibr" rid="B25">Konradt et&#xa0;al., 2011</xref>). Neutrophil migration requires dynamic reorganization of the actin cytoskeleton, as well as the membrane association of ERM family proteins acting as actin cytoskeletal linkers the hyaluronan receptor CD44 adhesion molecule. Critically, ERM protein association with the plasma membranes requires PIP<sub>2</sub> (<xref ref-type="bibr" rid="B25">Konradt et&#xa0;al., 2011</xref>). Through PIP<sub>2</sub> hydrolysis, IpgD prevents the membrane association of ERM proteins and neutrophil migration.</p>
</sec>
<sec id="s3">
<title>IpgD as a key regulator of global and local Ca<sup>2+</sup> signals</title>
<p>Because PI4,5P<sub>2</sub> is the main substrate utilized by PLCs to generate InsP<sub>3</sub>, IpgD also depresses InsP<sub>3</sub> levels by decreasing PI4,5P<sub>2</sub> levels. InsP<sub>3</sub> is the key second messenger leading to Ca<sup>2+</sup> release (<xref ref-type="bibr" rid="B53">Sun et&#xa0;al., 2017</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). During prolonged infection kinetics, IpgD virtually abolishes cytosolic Ca<sup>2+</sup> increase linked to release from internal stores. The inhibition of InsP<sub>3</sub>-mediated signaling is expected to have versatile implications during infection. Notably, cytosolic Ca<sup>2+</sup> increase is required for the activation of numerous Ca<sup>2+</sup> proteases (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). As a result of its inhibitory effects on Ca<sup>2+</sup> signaling, IpgD delays the Ca<sup>2+</sup>-dependent activation of calpains and disassembly of cell adhesion structures, thereby preserving the integrity of <italic>Shigella</italic>-infected cells (<xref ref-type="bibr" rid="B53">Sun et&#xa0;al., 2017</xref>).</p>
<p>Beyond inhibition, IpgD also has modulatory effects on <italic>Shigella</italic>-induced Ca<sup>2+</sup> signals at the early stages of bacterial invasion. Shortly upon contact with host cells, <italic>Shigella</italic> induces atypical Ca<sup>2+</sup> microdomains required for actin polymerization at invasion sites (<xref ref-type="bibr" rid="B55">Tran Van Nhieu et&#xa0;al., 2013</xref>). These <italic>Shigella</italic>-induced Ca<sup>2+</sup> microdomains correspond to local Ca<sup>2+</sup> increase, which duration reaching up to several seconds depend on the dense actin meshwork triggered by bacteria at entry sites and localized aspects of PLC activation and InsP<sub>3</sub> production at sites of bacteria-host cell membranes contact (<xref ref-type="bibr" rid="B55">Tran Van Nhieu et&#xa0;al., 2013</xref>). By limiting InsP<sub>3</sub> production, IpgD favors the formation of Ca<sup>2+</sup> microdomains at invasion sites, while limiting the elicitation of global Ca<sup>2+</sup> responses during the early stages of <italic>Shigella</italic> invasion (<xref ref-type="bibr" rid="B53">Sun et&#xa0;al., 2017</xref>).</p>
</sec>
<sec id="s4">
<title>Localized action of IpgD- stimulating actin polymerization during <italic>Shigella</italic> entry</title>
<p>During contact between <italic>Shigella</italic> and target epithelial cells, ruffles are formed locally in the near surrounding of the bacteria. While bacterial invasion was initially described to occur through macropinocytosis, advanced imaging showed that macropinosome formation and <italic>Shigella</italic> entry are two distinct processes (<xref ref-type="bibr" rid="B63">Weiner et&#xa0;al., 2016</xref>). Ruffling that is required for the generation of macropinosomes is driven by the Arp2/3-dependent polymerization of actin occurring downstream of activation of the small GTPase Rac (<xref ref-type="bibr" rid="B35">Mounier et&#xa0;al., 1999</xref>; <xref ref-type="bibr" rid="B11">Dum&#xe9;nil et&#xa0;al., 2000</xref>). Two <italic>Shigella</italic> type III effectors have been implicated in Rac activation and actin polymerization at invasion sites. The type III effector IpgB1 is a RacGEF amplifying actin polymerization and ruffle formation at a distance from the bacterial contact-site. The translocon component IpaC triggers actin polymerization at the intimate bacterial contact site by triggering the recruitment of the Src tyrosine kinase, to form an actin coat structure reminiscent of phagocytic cups in macrophages (<xref ref-type="bibr" rid="B56">Valencia-Gallardo et&#xa0;al., 2015</xref>). The mechanistic links between Src and Rac activation remain to be characterized and could involve phospholipases C and Ca<sup>2+</sup> signaling (<xref ref-type="bibr" rid="B55">Tran Van Nhieu et&#xa0;al., 2013</xref>).</p>
<p>The IpgD PIP<sub>2</sub> phosphatase activity favors actin polymerization at bacterial invasion sites. Seminal studies attributed this observation to the hydrolysis of PIP<sub>2</sub> leading to disassembly of cortical actin and stimulating <italic>de novo</italic> actin polymerization in close proximity to invading <italic>Shigella</italic> (<xref ref-type="bibr" rid="B37">Niebuhr et&#xa0;al., 2002</xref>). It became clear, however, that IpgD could regulate actin polymerization at invasion sites in various non-exclusive manners. Through PI5P production, IpgD may lead to the recruitment of the Rac GEF Tiam1 and Rac activation at membrane contacting bacteria (<xref ref-type="bibr" rid="B57">Viaud et&#xa0;al., 2014</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). Binding of PI5P to Tiam-1 stimulates its intrinsic GEF activity and Rac activation in ruffles and endosomes (<xref ref-type="bibr" rid="B57">Viaud et&#xa0;al., 2014</xref>). Also, IpgD was shown to promote the recruitment of the Arf6 GEF ARNO, likely <italic>via</italic> an increase of PI(3,4,5)P<sub>3</sub> levels insuring a feed-back loop of Arf6 activation required for efficient invasion (<xref ref-type="bibr" rid="B13">Garza-Mayers et&#xa0;al., 2015</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). While this was not investigated for <italic>Shigella</italic>, ARNO and Arf6 may potentiate actin polymerization in concert with the Rac-dependent recruitment of the Abi/WAVE complex as described for the <italic>Salmonella</italic> SopB ortholog (<xref ref-type="bibr" rid="B20">Humphreys et&#xa0;al., 2012</xref>; <xref ref-type="bibr" rid="B21">2013</xref>). Another interaction partner of IpgD could be the BAR domain protein TOCA-1 that also interacts with other <italic>Shigella</italic> effectors; nevertheless, the implication of this interaction requires more investigation (<xref ref-type="bibr" rid="B32">Miller et&#xa0;al., 2018</xref>).</p>
</sec>
<sec id="s5">
<title>IpgD-mediated regulation of actin after host cell entry</title>
<p>Upon completion of the bacterial invasion process, a thick, so-called &#x201c;cocoon&#x201d; forms <italic>de novo</italic> around the BCV (<xref ref-type="bibr" rid="B27">K&#xfc;hn et&#xa0;al., 2020</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>) This actin cocoon represents a unique structure implicating various cytoskeletal proteins and actin nucleation by the Arp2/3 complex. It is possible to speculate that through its N&#x3b5;-fatty acetylase activity the T3SS effector IcsB is required for persistent activity of the Arp2/3 complex at the actin cocoon, while the initial recruitment of this actin nucleating complex depends on the activation of the Cdc42 small GTPase, at the onset of bacterial invasion (<xref ref-type="bibr" rid="B29">Liu et&#xa0;al., 2018</xref>). The role of the actin cocoon needs to be further investigated, however evidence shows that it is required for the efficient removal of damaged vacuolar remnants following rupture, that would otherwise impair bacterial spreading. Also, the cocoon may prevent recognition of the BCV by the cell autonomous immune system. Intriguingly, IpgD appears also involved in actin cocoon regulation, however it remains to be investigated how IpgD-mediated actin polymerization/depolymerization functions are spatio-temporally regulated.</p>
<p>Following vacuolar escape, <italic>Shigella</italic> replicates freely in the cell cytosol and disseminate from cell-to-cell. Cell-to-cell spreading is permitted by the bacterial ability to use actin-based motility, pushing bacteria-containing protrusion into adjacent cells (<xref ref-type="bibr" rid="B60">Weddle and Agaisse, 2018</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). Upon lysis of the protrusion&#x2019;s double membrane, <italic>Shigella</italic> re-iterates replication and cell-to-cell spreading thereby disseminating over large area of the epithelial layer. Key to this process, engulfment of the bacteria-containing protrusions requires the disassembly of actin filaments at the protrusion&#x2019;s membranes. IpgD was shown to contribute to the down-regulation of actin structures in these protrusions, thus favoring their resolution into double-membrane containing secondary BCVs preceding bacterial escape into the cytosol within neighboring cells and bacteria dissemination (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). This highlights the diverse roles of IpgD during successive steps of <italic>Shigella</italic> invasion (<xref ref-type="bibr" rid="B26">K&#xf6;seo&#x11f;lu et&#xa0;al., 2022</xref>).</p>
</sec>
<sec id="s6">
<title>IpgD requirement during the events of vacuolar rupture</title>
<p>The BCV that englobes <italic>Shigella</italic> during its uptake differs from the macropinocytic vesicles in its close proximity, reinforcing differences between <italic>Shigella</italic> invasion and canonical macropinocytosis (<xref ref-type="bibr" rid="B31">Mellouk et&#xa0;al., 2014</xref>; <xref ref-type="bibr" rid="B63">Weiner et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B7">Chang et&#xa0;al., 2021</xref>). While bacteria invasion triggers the formation of ruffles and exocytic vesicles, the nascent BCV is formed in tight apposition with the bacterial body. The duality of macropinosome and BCV formation has been discovered through large volume correlative light and electron microscopy (CLEM) studies (<xref ref-type="bibr" rid="B63">Weiner et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B62">Weiner and Enninga, 2019</xref>). The newly formed macropinosomes cluster around the BCV within minutes upon bacterial invasion, however fusion between both compartments could not be observed. Furthermore, breaching of the BCV membrane takes place precisely at the time when it contacts macropinosomes. Macropinosomes-BCV contact sites can be visualized by 3D ultrastructural techniques, such as focused ion beam scanning EM. Studies on the formation of both compartments indicate that IpgD plays a main role in efficient vacuolar disruption (<xref ref-type="bibr" rid="B31">Mellouk et&#xa0;al., 2014</xref>; <xref ref-type="bibr" rid="B8">Chang et&#xa0;al., 2020</xref>). <italic>ipgD</italic> mutants show similar internalization kinetics as WT <italic>Shigella</italic>, however, the usually observed massive damage of the BCV is delayed. The differences between simple BCV damage and the unwrapping of the damaged BCV leaflets can be measured combining two microscopic assays; fluorescently tagged galectins that get recruited to the inner leaflet of the BCV upon its damage show the morphology of BCV disassembly, while a sensitive enzymatic FRET reporter provides precise information on the initial damage of BCV damage. Two clear defects in relation to vacuolar rupture can be attributed to the <italic>ipgD</italic> mutant <italic>Shigella</italic> strain: i, the absence of macropinosomes at bacterial entry sites; ii, the incapacity of invading <italic>Shigella</italic> to strip off BCV remnants that are typically transported several micrometers away upon BCV rupture. Potentially, the bacteria entrapped in broken BCVs are readily targeted by the xenophagy machinery. During these steps, forming macropinosomes become coated with Rab11, Rab8 and the exocyst complex (<xref ref-type="bibr" rid="B8">Chang et&#xa0;al., 2020</xref>), as shown by proteomics analysis of macropinosomes isolated from epithelial cells infected with <italic>Shigella</italic> (<xref ref-type="bibr" rid="B8">Chang et&#xa0;al., 2020</xref>; <xref ref-type="bibr" rid="B52">St&#xe9;venin et&#xa0;al., 2021</xref>) (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). The resuting Rab/exocyst coating is required for targeting these compartments to the BCV before its rupture. Upon Rab11 knockdown, depletion of key constituents of the exocyst complex, such as Exo70, or by using interfering peptides that hamper exocyst complex formation, the BCV and macropinosomes form less contacts. Also, damaged BCV membranes remain in close apposition to the bacteria (<xref ref-type="bibr" rid="B31">Mellouk et&#xa0;al., 2014</xref>; <xref ref-type="bibr" rid="B8">Chang et&#xa0;al., 2020</xref>), impairing intra- and inter-cellular bacterial spread as observed for a <italic>Shigella ipgD</italic> mutant. Whether IpgD acts only on the surrounding macropinosomes, on the BCV, or simultaneously on both compartments will require further investigation.</p>
</sec>
<sec id="s7">
<title>IpgD orthologs: The role of SopB during <italic>Salmonella</italic> invasion</title>
<p>
<italic>Salmonella</italic> SopB (or SigD) was already shown to be an ortholog of IpgD more than two decades ago and it was further characterized as PI phosphatase (<xref ref-type="bibr" rid="B38">Norris et&#xa0;al., 1998</xref>). Both proteins exhibit an unstructured N-terminus likely involved in chaperone-binding and secretion by the T3SS, as well as a P-loop containing a Cys-X5-Arg motif that is a signature of the active site of Mg<sup>2+</sup>-independent phosphatases (<xref ref-type="bibr" rid="B38">Norris et&#xa0;al., 1998</xref>; <xref ref-type="bibr" rid="B37">Niebuhr et&#xa0;al., 2002</xref>) (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2A</bold>
</xref>, red box). In addition, a discrete hydrophobic region involved in membrane interaction has been described for SopB and suggested for IpgD (<xref ref-type="bibr" rid="B39">Patel et&#xa0;al., 2009</xref>; <xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2A</bold>
</xref>, yellow box MIM1. Using <italic>in silico</italic> computational methods for predicting topology of transmembranes 3D structures (DAS-TMpred and DeepEMPred), we confirmed MIM1 and identified a longer MIM2 for IpgD (blue box) (<xref ref-type="bibr" rid="B10">Cserz&#xf6; et&#xa0;al., 1997</xref>; <xref ref-type="bibr" rid="B19">Hsu and Mao, 2015</xref>; <xref ref-type="bibr" rid="B61">Weigele et&#xa0;al., 2017</xref>; <xref ref-type="bibr" rid="B59">Wang et&#xa0;al., 2022</xref>). We also predicted a similar MIM2 in SopB through its sequence and structural concordances (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2A</bold>
</xref>, blue box MIM2), that includes a motif conserved in host cell phosphatases formerly identified by <xref ref-type="bibr" rid="B38">Norris et&#xa0;al., 1998</xref>. (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2A</bold>
</xref>; brown empty box)). Using Pymol, we generated an <italic>in silico</italic> model of IpgD that shows the spatial proximity of both MIMs to the P-loop (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2B</bold>
</xref>, left). This organization is similar to that of host phosphoinositides phosphatases, where MIMs are observed in close proximity to the active site and are presumed to facilitate accessibility to the phosphoinositide following membrane insertion (<xref ref-type="bibr" rid="B19">Hsu and Mao, 2015</xref>). Further stressing similarities with host lipid phosphatases, the IpgD surface surrounding the catalytic P-loop region shows a high density of positive charges likely involved in interaction with the negatively charged membranes embedding the phosphoinositide substrate (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2B</bold>
</xref>). Alphafold structure simulations of IpgD and SopB demonstrate their high levels of homology with an identical organization of MIMs and conserved residues surrounding the P-loop region (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2C</bold>
</xref>, yellow, cyan and red). Together, this analysis also shows conserved structural features and domain organization between IpgD, SopB and phosphoinosited phosphatases (<xref ref-type="bibr" rid="B19">Hsu and Mao, 2015</xref>). Functionally however, while <italic>in vitro</italic>, IpgD preferentially targets for PI(4,5)P<sub>2,</sub> SopB shows preferential activity towards PI(3,4)P<sub>2.</sub> How the <italic>in vitro</italic> SopB phosphatase activity towards PI(3,4)P<sub>2</sub> relates to the increased PI(3,4)P<sub>2</sub> levels associated with the SopB phosphotransferase activity has remained unclear until recently, when SopB was identified as phosphotransferase similar to IpgD (<xref ref-type="bibr" rid="B58">Walpole et&#xa0;al., 2022</xref>) (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2</bold>
</xref>). During host cell challenge with <italic>Salmonella</italic>, SopB has been found to increase PI3P levels, which sets it apart from the production of PI5P in the case of IpgD (<xref ref-type="bibr" rid="B18">Hernandez et&#xa0;al., 2004</xref>). As for IpgD, SopB activates Akt but SopB-mediated Akt activation occurs through a process that requires PI(3,4)P<sub>2</sub> and is Wortmannin insensitive (<xref ref-type="bibr" rid="B50">Steele-Mortimer et&#xa0;al., 2000</xref>; <xref ref-type="bibr" rid="B9">Cooper et&#xa0;al., 2011</xref>). Local targeting of SopB at the onset of these events has been shown to depend on myosin 6 tethering to the entry site <italic>via</italic> SopE (<xref ref-type="bibr" rid="B4">Brooks et&#xa0;al., 2017</xref>). Akt activation promotes intracellular bacterial growth, modulates the immune response, enhances M cell growth within the intestinal epithelium, and potentially plays a role in infection associated carcinoma (<xref ref-type="bibr" rid="B24">Knodler et&#xa0;al., 2005</xref>; <xref ref-type="bibr" rid="B28">Kum et&#xa0;al., 2011</xref>; <xref ref-type="bibr" rid="B54">Tahoun et&#xa0;al., 2012</xref>; <xref ref-type="bibr" rid="B48">Scanu et&#xa0;al., 2015</xref>). More recently, SopB has also been linked with the intracellular niche formation of <italic>Salmonella</italic> during epithelial infection (<xref ref-type="bibr" rid="B52">St&#xe9;venin et&#xa0;al., 2021</xref>). SopB reprograms phosphoinositides at plasma membranes of <italic>Salmonella</italic> invasion sites (<xref ref-type="bibr" rid="B18">Hernandez et&#xa0;al., 2004</xref>; <xref ref-type="bibr" rid="B42">Piscatelli et&#xa0;al., 2016</xref>). Furthermore, SopB plays a major role in extracting membranes from the BCV during the early invasion phase through the formation of SNX3-positive tubules (<xref ref-type="bibr" rid="B5">Bujny et&#xa0;al., 2008</xref>; <xref ref-type="bibr" rid="B3">Braun et&#xa0;al., 2010</xref>).</p>
<fig id="f2" position="float">
<label>Figure&#xa0;2</label>
<caption>
<p>Conserved phosphoinositide phosphatase features of <italic>Shigella</italic> IpgD and <italic>Salmonella</italic> SopB. <bold>(A)</bold> Sequence alignments of MIMs and P-loop motives in IpgD and SopB. Bold characters: conserved residues. Yellow box: MIM1, blue box: MIM2, red box: P-loop. The MIMs were identified experimentally and through computational learning, approaches (see text). <bold>(B)</bold> Left: Alphafold-based <italic>in silico</italic> 3D structure analysis of IpgD (<xref ref-type="bibr" rid="B23">Jumper et&#xa0;al., 2021</xref>) followed by PyMOL molecular visualization (<xref ref-type="bibr" rid="B49">Schr&#xf6;dinger and DeLano 2020</xref>) showing the P-loop and surrounding MIMs. Right: surface charge distribution. Blue: positive charges, red: negative charges. <bold>(C)</bold> High structural homologies between IpgD and SopB. Green: IpgD, purple: SopB, yellow: MIM1, cyan: MIM2, red: P-loop, black: conserved aminoacids. Left and Middle: Alphafold-based 3D structures of IpgD and SopB. Right: overlaid 3D structures showing their high homology. <bold>(D)</bold> Enzymatic reactions of IpgD and SopB. PI(4,5)P<sub>2</sub> dephosphorylation either yields PI5P for both enzymes or PI3P in the case of SopB (left). The recently described phosphotransferase activity leads to PI(3,4)P<sub>2</sub> production that could be further dephosphorylated in PI3P (right) (<xref ref-type="bibr" rid="B58">Walpole et&#xa0;al., 2022</xref>).</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="fcimb-12-1012533-g002.tif"/>
</fig>
<p>Contrasting with <italic>Shigella</italic> invasion, <italic>Salmonella</italic>-induced macropinosomes can fuse with the BCV, a process that is SopB -independent. SopB appears rather involved in regulating the stability of the BCV, however through the regulation of membrane influx and efflux to and from the BCV (<xref ref-type="bibr" rid="B51">St&#xe9;venin et&#xa0;al., 2019</xref>). Therefore, despite the similarities of the enzymatic function of the <italic>Shigella</italic> and <italic>Salmonella</italic> effectors IpgD and SopB, their mechanistic involvement in controlling the intracellular niche is distinct for both pathogens, perhaps due to differences in the composition of the macropinosomes forming around the respective BCVs.</p>
</sec>
<sec id="s8">
<title>Perspectives</title>
<p>The function of IpgD during <italic>Shigella</italic> has been studied for more than two decades and has revealed its crucial involvement during entry, intracellular niche formation, dissemination as well as host immune signaling. More generally, this reflects the importance of phosphoinositide reprogramming during the course of <italic>Shigella</italic> infection. Phosphoinositides are key components linking membrane remodeling, cytoskeletal rearrangements, and complex signaling pathways including Ca<sup>2+</sup>. Based on recent discoveries reviewed here, more work is required to obtain a comprehensive understanding about the role of IpgD for <italic>Shigella</italic> infection. It will be particularly interesting to integrate the IpgD phosphotransferase activity with regards to its largely described phosphatase activity. As PI(3,4)P<sub>2</sub> and PI(5)P trigger different signals, it will be important to decipher what is the relevance for each of the different activities during cell infection by <italic>Shigella</italic>. A comprehensive analysis of the similarities and differences between the <italic>Shigella</italic> effector IpgD and SopB of <italic>Salmonella</italic> will also be useful to understand the differences of their respective intracellular lifestyles. In this regard, a detailed structure-function analysis targeting specific domains of these exciting effectors will be highly informative. Novel in silico approaches, such as Alphafold are powerful tools to achieve this (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2</bold>
</xref>). Also, as IpgD acts locally at the entry site as well as globally within targeted cells, including at sites of cell-to-cell spread, it will be important to better understand the spatiotemporal organization of the action of IpgD. In this context, it would be intriguing to investigate whether IpgD gets post-translationally modified upon translocation into host cells as has been suggested for SopB (<xref ref-type="bibr" rid="B39">Patel et&#xa0;al., 2009</xref>; <xref ref-type="bibr" rid="B44">Popa et&#xa0;al., 2016</xref>). Finally, model tissues through organ-on-a-chip devices or co-cultures (<xref ref-type="bibr" rid="B14">Grassart et&#xa0;al., 2019</xref>; <xref ref-type="bibr" rid="B33">Mitchell et&#xa0;al., 2020</xref>; <xref ref-type="bibr" rid="B47">Rey et&#xa0;al., 2020</xref>) may provide with important insights to confirm the role of IpgD in <italic>Shigella</italic> pathophysiology.</p>
</sec>
<sec id="s9" sec-type="author-contributions">
<title>Author contributions</title>
<p>GTVN, JE, and PL-L contributed to conception and design of the study. GTVN, JE wrote the first draft of the manuscript. PL-L organized 3D structures and analysis. GTVN, JE, PL-L wrote sections of the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version.</p>
</sec>
<sec id="s10" sec-type="funding-information">
<title>Funding</title>
<p>JE and GTVN are funded by the Agence National pour la Recherche Projects &#x201c;PureMagRupture&#x201d; (JE, GTVN) and &#x201c;RabReprogram&#x201d; (JE).</p>
</sec>
<sec id="s11" sec-type="acknowledgement">
<title>Acknowledgments</title>
<p>JE acknowledges support from the Institut Pasteur and the European Research Council (ERC-CoG &#x201c;Endosubvert&#x201d;). JE is member of the LabEx IBEID and Milieu Interieur. GTVN is supported by the Inserm (Institut National de la Sant&#xe9; et de la Recherche M&#xe9; dicale) and the CNRS (Centre National de la Recherche Scientifique)</p>
</sec>
<sec id="s12" sec-type="COI-statement">
<title>Conflict of interest</title>
<p>The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.</p>
</sec>
<sec id="s13" sec-type="disclaimer">
<title>Publisher&#x2019;s note</title>
<p>All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.</p>
</sec>
</body>
<back>
<ref-list>
<title>References</title>
<ref id="B1">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Boal</surname> <given-names>F.</given-names>
</name>
<name>
<surname>Mansour</surname> <given-names>R.</given-names>
</name>
<name>
<surname>Gayral</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Saland</surname> <given-names>E.</given-names>
</name>
<name>
<surname>Chicanne</surname> <given-names>G.</given-names>
</name>
<name>
<surname>Xuereb</surname> <given-names>J.-M.</given-names>
</name>
<etal/>
</person-group>. (<year>2015</year>). <article-title>TOM1 is a PI5P effector involved in the regulation of endosomal maturation</article-title>. <source>J. Cell Sci.</source> <volume>128</volume>, <fpage>815</fpage>&#x2013;<lpage>827</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1242/jcs.166314</pub-id>
</citation>
</ref>
<ref id="B2">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Boal</surname> <given-names>F.</given-names>
</name>
<name>
<surname>Puhar</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Xuereb</surname> <given-names>J.-M.</given-names>
</name>
<name>
<surname>Kunduzova</surname> <given-names>O.</given-names>
</name>
<name>
<surname>Sansonetti</surname> <given-names>P. J.</given-names>
</name>
<name>
<surname>Payrastre</surname> <given-names>B.</given-names>
</name>
<etal/>
</person-group>. (<year>2016</year>). <article-title>PI5P triggers ICAM-1 degradation in shigella infected cells, thus dampening immune cell recruitment</article-title>. <source>Cell Rep.</source> <volume>14</volume>, <fpage>750</fpage>&#x2013;<lpage>759</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.celrep.2015.12.079</pub-id>
</citation>
</ref>
<ref id="B3">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Braun</surname> <given-names>V.</given-names>
</name>
<name>
<surname>Wong</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Landekic</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Hong</surname> <given-names>W. J.</given-names>
</name>
<name>
<surname>Grinstein</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Brumell</surname> <given-names>J. H.</given-names>
</name>
</person-group> (<year>2010</year>). <article-title>Sorting nexin 3 (SNX3) is a component of a tubular endosomal network induced by salmonella and involved in maturation of the salmonella-containing vacuole</article-title>. <source>Cell Microbiol.</source> <volume>12</volume>, <fpage>1352</fpage>&#x2013;<lpage>1367</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1111/j.1462-5822.2010.01476.x</pub-id>
</citation>
</ref>
<ref id="B4">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Brooks</surname> <given-names>A. B. E.</given-names>
</name>
<name>
<surname>Humphreys</surname> <given-names>D.</given-names>
</name>
<name>
<surname>Singh</surname> <given-names>V.</given-names>
</name>
<name>
<surname>Davidson</surname> <given-names>A. C.</given-names>
</name>
<name>
<surname>Arden</surname> <given-names>S. D.</given-names>
</name>
<name>
<surname>Buss</surname> <given-names>F.</given-names>
</name>
<etal/>
</person-group>. (<year>2017</year>). <article-title>MYO6 is targeted by salmonella virulence effectors to trigger PI3-kinase signaling and pathogen invasion into host cells</article-title>. <source>Proc. Natl. Acad. Sci. U.S.A.</source> <volume>114</volume>, <fpage>3915</fpage>&#x2013;<lpage>3920</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1073/pnas.1616418114</pub-id>
</citation>
</ref>
<ref id="B5">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Bujny</surname> <given-names>M. V.</given-names>
</name>
<name>
<surname>Ewels</surname> <given-names>P. A.</given-names>
</name>
<name>
<surname>Humphrey</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Attar</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Jepson</surname> <given-names>M. A.</given-names>
</name>
<name>
<surname>Cullen</surname> <given-names>P. J.</given-names>
</name>
</person-group> (<year>2008</year>). <article-title>Sorting nexin-1 defines an early phase of salmonella-containing vacuole-remodeling during salmonella infection</article-title>. <source>J. Cell Sci.</source> <volume>121</volume>, <fpage>2027</fpage>&#x2013;<lpage>2036</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1242/jcs.018432</pub-id>
</citation>
</ref>
<ref id="B6">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Burnaevskiy</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Fox</surname> <given-names>T. G.</given-names>
</name>
<name>
<surname>Plymire</surname> <given-names>D. A.</given-names>
</name>
<name>
<surname>Ertelt</surname> <given-names>J. M.</given-names>
</name>
<name>
<surname>Weigele</surname> <given-names>B. A.</given-names>
</name>
<name>
<surname>Selyunin</surname> <given-names>A. S.</given-names>
</name>
<etal/>
</person-group>. (<year>2013</year>). <article-title>Proteolytic elimination of n-myristoyl modifications by the shigella virulence factor IpaJ</article-title>. <source>Nature</source> <volume>496</volume>, <fpage>106</fpage>&#x2013;<lpage>109</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/nature12004</pub-id>
</citation>
</ref>
<ref id="B7">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Chang</surname> <given-names>Y.-Y.</given-names>
</name>
<name>
<surname>Enninga</surname> <given-names>J.</given-names>
</name>
<name>
<surname>St&#xe9;venin</surname> <given-names>V.</given-names>
</name>
</person-group> (<year>2021</year>). <article-title>New methods to decrypt emerging macropinosome functions during the host-pathogen crosstalk</article-title>. <source>Cell Microbiol.</source> <volume>23</volume>, <elocation-id>e13342</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.1111/cmi.13342</pub-id>
</citation>
</ref>
<ref id="B8">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Chang</surname> <given-names>Y.-Y.</given-names>
</name>
<name>
<surname>St&#xe9;venin</surname> <given-names>V.</given-names>
</name>
<name>
<surname>Duchateau</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Giai Gianetto</surname> <given-names>Q.</given-names>
</name>
<name>
<surname>Hourdel</surname> <given-names>V.</given-names>
</name>
<name>
<surname>Rodrigues</surname> <given-names>C. D.</given-names>
</name>
<etal/>
</person-group>. (<year>2020</year>). <article-title>Shigella hijacks the exocyst to cluster macropinosomes for efficient vacuolar escape</article-title>. <source>PloS Pathog.</source> <volume>16</volume>, <elocation-id>e1008822</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.1371/journal.ppat.1008822</pub-id>
</citation>
</ref>
<ref id="B9">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Cooper</surname> <given-names>K. G.</given-names>
</name>
<name>
<surname>Winfree</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Malik-Kale</surname> <given-names>P.</given-names>
</name>
<name>
<surname>Jolly</surname> <given-names>C.</given-names>
</name>
<name>
<surname>Ireland</surname> <given-names>R.</given-names>
</name>
<name>
<surname>Knodler</surname> <given-names>L. A.</given-names>
</name>
<etal/>
</person-group>. (<year>2011</year>). <article-title>Activation of akt by the bacterial inositol phosphatase, SopB, is wortmannin insensitive</article-title>. <source>PloS One</source> <volume>6</volume>, <elocation-id>e22260</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.1371/journal.pone.0022260</pub-id>
</citation>
</ref>
<ref id="B10">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Cserz&#xf6;</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Wallin</surname> <given-names>E.</given-names>
</name>
<name>
<surname>Simon</surname> <given-names>I.</given-names>
</name>
<name>
<surname>von Heijne</surname> <given-names>G.</given-names>
</name>
<name>
<surname>Elofsson</surname> <given-names>A.</given-names>
</name>
</person-group> (<year>1997</year>). <article-title>Prediction of transmembrane alpha-helices in prokaryotic membrane proteins: The dense alignment surface method</article-title>. <source>Protein Eng.</source> <volume>10</volume>, <fpage>673</fpage>&#x2013;<lpage>676</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1093/protein/10.6.673</pub-id>
</citation>
</ref>
<ref id="B11">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Dum&#xe9;nil</surname> <given-names>G.</given-names>
</name>
<name>
<surname>Sansonetti</surname> <given-names>P.</given-names>
</name>
<name>
<surname>Tran Van Nhieu</surname> <given-names>G.</given-names>
</name>
</person-group> (<year>2000</year>). <article-title>Src tyrosine kinase activity down-regulates rho-dependent responses during shigella entry into epithelial cells and stress fibre formation</article-title>. <source>J. Cell Sci.</source> <volume>113</volume> (<issue>Pt 1</issue>), <fpage>71</fpage>&#x2013;<lpage>80</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1242/jcs.113.1.71</pub-id>
</citation>
</ref>
<ref id="B12">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Enninga</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Mounier</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Sansonetti</surname> <given-names>P.</given-names>
</name>
<name>
<surname>Tran Van Nhieu</surname> <given-names>G.</given-names>
</name>
</person-group> (<year>2005</year>). <article-title>Secretion of type III effectors into host cells in real time</article-title>. <source>Nat. Methods</source> <volume>2</volume>, <fpage>959</fpage>&#x2013;<lpage>965</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/nmeth804</pub-id>
</citation>
</ref>
<ref id="B13">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Garza-Mayers</surname> <given-names>A. C.</given-names>
</name>
<name>
<surname>Miller</surname> <given-names>K. A.</given-names>
</name>
<name>
<surname>Russo</surname> <given-names>B. C.</given-names>
</name>
<name>
<surname>Nagda</surname> <given-names>D. V.</given-names>
</name>
<name>
<surname>Goldberg</surname> <given-names>M. B.</given-names>
</name>
</person-group> (<year>2015</year>). <article-title>Shigella flexneri regulation of ARF6 activation during bacterial entry <italic>via</italic> an IpgD-mediated positive feedback loop</article-title>. <source>mBio</source> <volume>6</volume>, <elocation-id>e02584-14</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.1128/mBio.02584-14</pub-id>
</citation>
</ref>
<ref id="B14">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Grassart</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Malard&#xe9;</surname> <given-names>V.</given-names>
</name>
<name>
<surname>Gobaa</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Sartori-Rupp</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Kerns</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Karalis</surname> <given-names>K.</given-names>
</name>
<etal/>
</person-group>. (<year>2019</year>). <article-title>Bioengineered human organ-on-Chip reveals intestinal microenvironment and mechanical forces impacting shigella infection</article-title>. <source>Cell Host Microbe</source> <volume>26</volume>, <fpage>435</fpage>&#x2013;<lpage>444.e4</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.chom.2019.08.007</pub-id>
</citation>
</ref>
<ref id="B15">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Hachani</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Biskri</surname> <given-names>L.</given-names>
</name>
<name>
<surname>Rossi</surname> <given-names>G.</given-names>
</name>
<name>
<surname>Marty</surname> <given-names>A.</given-names>
</name>
<name>
<surname>M&#xe9;nard</surname> <given-names>R.</given-names>
</name>
<name>
<surname>Sansonetti</surname> <given-names>P.</given-names>
</name>
<etal/>
</person-group>. (<year>2008</year>). <article-title>IpgB1 and IpgB2, two homologous effectors secreted <italic>via</italic> the mxi-spa type III secretion apparatus, cooperate to mediate polarized cell invasion and inflammatory potential of shigella flexenri</article-title>. <source>Microbes Infect.</source> <volume>10</volume>, <fpage>260</fpage>&#x2013;<lpage>268</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.micinf.2007.11.011</pub-id>
</citation>
</ref>
<ref id="B16">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Handa</surname> <given-names>Y.</given-names>
</name>
<name>
<surname>Suzuki</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Ohya</surname> <given-names>K.</given-names>
</name>
<name>
<surname>Iwai</surname> <given-names>H.</given-names>
</name>
<name>
<surname>Ishijima</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Koleske</surname> <given-names>A. J.</given-names>
</name>
<etal/>
</person-group>. (<year>2007</year>). <article-title>Shigella IpgB1 promotes bacterial entry through the ELMO-Dock180 machinery</article-title>. <source>Nat. Cell Biol.</source> <volume>9</volume>, <fpage>121</fpage>&#x2013;<lpage>128</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/ncb1526</pub-id>
</citation>
</ref>
<ref id="B17">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Heindl</surname> <given-names>J. E.</given-names>
</name>
<name>
<surname>Saran</surname> <given-names>I.</given-names>
</name>
<name>
<surname>Yi</surname> <given-names>C.</given-names>
</name>
<name>
<surname>Lesser</surname> <given-names>C. F.</given-names>
</name>
<name>
<surname>Goldberg</surname> <given-names>M. B.</given-names>
</name>
</person-group> (<year>2010</year>). <article-title>Requirement for formin-induced actin polymerization during spread of shigella flexneri</article-title>. <source>Infect. Immun.</source> <volume>78</volume>, <fpage>193</fpage>&#x2013;<lpage>203</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1128/IAI.00252-09</pub-id>
</citation>
</ref>
<ref id="B18">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Hernandez</surname> <given-names>L. D.</given-names>
</name>
<name>
<surname>Hueffer</surname> <given-names>K.</given-names>
</name>
<name>
<surname>Wenk</surname> <given-names>M. R.</given-names>
</name>
<name>
<surname>Gal&#xe1;n</surname> <given-names>J. E.</given-names>
</name>
</person-group> (<year>2004</year>). <article-title>Salmonella modulates vesicular traffic by altering phosphoinositide metabolism</article-title>. <source>Science</source> <volume>304</volume>, <fpage>1805</fpage>&#x2013;<lpage>1807</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1126/science.1098188</pub-id>
</citation>
</ref>
<ref id="B19">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Hsu</surname> <given-names>F.</given-names>
</name>
<name>
<surname>Mao</surname> <given-names>Y.</given-names>
</name>
</person-group> (<year>2015</year>). <article-title>The structure of phosphoinositide phosphatases: Insights into substrate specificity and catalysis</article-title>. <source>Biochim. Biophys. Acta</source> <volume>1851</volume>, <fpage>698</fpage>&#x2013;<lpage>710</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.bbalip.2014.09.015</pub-id>
</citation>
</ref>
<ref id="B20">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Humphreys</surname> <given-names>D.</given-names>
</name>
<name>
<surname>Davidson</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Hume</surname> <given-names>P. J.</given-names>
</name>
<name>
<surname>Koronakis</surname> <given-names>V.</given-names>
</name>
</person-group> (<year>2012</year>). <article-title>Salmonella virulence effector SopE and host GEF ARNO cooperate to recruit and activate WAVE to trigger bacterial invasion</article-title>. <source>Cell Host Microbe</source> <volume>11</volume>, <fpage>129</fpage>&#x2013;<lpage>139</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.chom.2012.01.006</pub-id>
</citation>
</ref>
<ref id="B21">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Humphreys</surname> <given-names>D.</given-names>
</name>
<name>
<surname>Davidson</surname> <given-names>A. C.</given-names>
</name>
<name>
<surname>Hume</surname> <given-names>P. J.</given-names>
</name>
<name>
<surname>Makin</surname> <given-names>L. E.</given-names>
</name>
<name>
<surname>Koronakis</surname> <given-names>V.</given-names>
</name>
</person-group> (<year>2013</year>). <article-title>Arf6 coordinates actin assembly through the WAVE complex, a mechanism usurped by salmonella to invade host cells</article-title>. <source>Proc. Natl. Acad. Sci. U.S.A.</source> <volume>110</volume>, <fpage>16880</fpage>&#x2013;<lpage>16885</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1073/pnas.1311680110</pub-id>
</citation>
</ref>
<ref id="B22">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Jennings</surname> <given-names>E.</given-names>
</name>
<name>
<surname>Thurston</surname> <given-names>T. L. M.</given-names>
</name>
<name>
<surname>Holden</surname> <given-names>D. W.</given-names>
</name>
</person-group> (<year>2017</year>). <article-title>Salmonella SPI-2 type III secretion system effectors: Molecular mechanisms and physiological consequences</article-title>. <source>Cell Host Microbe</source> <volume>22</volume>, <fpage>217</fpage>&#x2013;<lpage>231</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.chom.2017.07.009</pub-id>
</citation>
</ref>
<ref id="B23">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Jumper</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Evans</surname> <given-names>R.</given-names>
</name>
<name>
<surname>Pritzel</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Green</surname> <given-names>T.</given-names>
</name>
<name>
<surname>Figurnov</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Ronneberger</surname> <given-names>O.</given-names>
</name>
<etal/>
</person-group>. (<year>2021</year>). <article-title>Highly accurate protein structure prediction with AlphaFold</article-title>. <source>Nature</source> <volume>596</volume>, <fpage>583</fpage>&#x2013;<lpage>589</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/s41586-021-03819-2</pub-id>
</citation>
</ref>
<ref id="B24">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Knodler</surname> <given-names>L. A.</given-names>
</name>
<name>
<surname>Finlay</surname> <given-names>B. B.</given-names>
</name>
<name>
<surname>Steele-Mortimer</surname> <given-names>O.</given-names>
</name>
</person-group> (<year>2005</year>). <article-title>The salmonella effector protein SopB protects epithelial cells from apoptosis by sustained activation of akt</article-title>. <source>J. Biol. Chem.</source> <volume>280</volume>, <fpage>9058</fpage>&#x2013;<lpage>9064</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1074/jbc.M412588200</pub-id>
</citation>
</ref>
<ref id="B25">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Konradt</surname> <given-names>C.</given-names>
</name>
<name>
<surname>Frigimelica</surname> <given-names>E.</given-names>
</name>
<name>
<surname>Nothelfer</surname> <given-names>K.</given-names>
</name>
<name>
<surname>Puhar</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Salgado-Pabon</surname> <given-names>W.</given-names>
</name>
<name>
<surname>di Bartolo</surname> <given-names>V.</given-names>
</name>
<etal/>
</person-group>. (<year>2011</year>). <article-title>The shigella flexneri type three secretion system effector IpgD inhibits T cell migration by manipulating host phosphoinositide metabolism</article-title>. <source>Cell Host Microbe</source> <volume>9</volume>, <fpage>263</fpage>&#x2013;<lpage>272</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.chom.2011.03.010</pub-id>
</citation>
</ref>
<ref id="B26">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>K&#xf6;seo&#x11f;lu</surname> <given-names>V. K.</given-names>
</name>
<name>
<surname>Jones</surname> <given-names>M. K.</given-names>
</name>
<name>
<surname>Agaisse</surname> <given-names>H.</given-names>
</name>
</person-group> (<year>2022</year>). <article-title>The type 3 secretion effector IpgD promotes s. flexneri dissemination</article-title>. <source>PloS Pathog.</source> <volume>18</volume>, <elocation-id>e1010324</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.1371/journal.ppat.1010324</pub-id>
</citation>
</ref>
<ref id="B27">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>K&#xfc;hn</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Bergqvist</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Gil</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Valenzuela</surname> <given-names>C.</given-names>
</name>
<name>
<surname>Barrio</surname> <given-names>L.</given-names>
</name>
<name>
<surname>Lebreton</surname> <given-names>S.</given-names>
</name>
<etal/>
</person-group>. (<year>2020</year>). <article-title>Actin assembly around the shigella-containing vacuole promotes successful infection</article-title>. <source>Cell Rep.</source> <volume>31</volume>, <elocation-id>107638</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.celrep.2020.107638</pub-id>
</citation>
</ref>
<ref id="B28">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Kum</surname> <given-names>W. W. S.</given-names>
</name>
<name>
<surname>Lo</surname> <given-names>B. C.</given-names>
</name>
<name>
<surname>Yu</surname> <given-names>H. B.</given-names>
</name>
<name>
<surname>Finlay</surname> <given-names>B. B.</given-names>
</name>
</person-group> (<year>2011</year>). <article-title>Protective role of Akt2 in salmonella enterica serovar typhimurium-induced gastroenterocolitis</article-title>. <source>Infect. Immun.</source> <volume>79</volume>, <fpage>2554</fpage>&#x2013;<lpage>2566</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1128/IAI.01235-10</pub-id>
</citation>
</ref>
<ref id="B29">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Liu</surname> <given-names>W.</given-names>
</name>
<name>
<surname>Zhou</surname> <given-names>Y.</given-names>
</name>
<name>
<surname>Peng</surname> <given-names>T.</given-names>
</name>
<name>
<surname>Zhou</surname> <given-names>P.</given-names>
</name>
<name>
<surname>Ding</surname> <given-names>X.</given-names>
</name>
<name>
<surname>Li</surname> <given-names>Z.</given-names>
</name>
<etal/>
</person-group>. (<year>2018</year>). <article-title>N&#x3f5;-fatty acylation of multiple membrane-associated proteins by shigella IcsB effector to modulate host function</article-title>. <source>Nat. Microbiol.</source> <volume>3</volume>, <fpage>996</fpage>&#x2013;<lpage>1009</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/s41564-018-0215-6</pub-id>
</citation>
</ref>
<ref id="B30">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>L&#xf3;pez-Montero</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Enninga</surname> <given-names>J.</given-names>
</name>
</person-group> (<year>2018</year>). <article-title>Diverted recycling-shigella subversion of rabs</article-title>. <source>Small GTPases</source> <volume>9</volume>, <fpage>365</fpage>&#x2013;<lpage>374</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1080/21541248.2016.1240494</pub-id>
</citation>
</ref>
<ref id="B31">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Mellouk</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Weiner</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Aulner</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Schmitt</surname> <given-names>C.</given-names>
</name>
<name>
<surname>Elbaum</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Shorte</surname> <given-names>S. L.</given-names>
</name>
<etal/>
</person-group>. (<year>2014</year>). <article-title>Shigella subverts the host recycling compartment to rupture its vacuole</article-title>. <source>Cell Host Microbe</source> <volume>16</volume>, <fpage>517</fpage>&#x2013;<lpage>530</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.chom.2014.09.005</pub-id>
</citation>
</ref>
<ref id="B32">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Miller</surname> <given-names>K. A.</given-names>
</name>
<name>
<surname>Garza-Mayers</surname> <given-names>A. C.</given-names>
</name>
<name>
<surname>Leung</surname> <given-names>Y.</given-names>
</name>
<name>
<surname>Goldberg</surname> <given-names>M. B.</given-names>
</name>
</person-group> (<year>2018</year>). <article-title>Identification of interactions among host and bacterial proteins and evaluation of their role early during shigella flexneri infection</article-title>. <source>Microbiol. (Reading)</source> <volume>164</volume>, <fpage>540</fpage>&#x2013;<lpage>550</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1099/mic.0.000637</pub-id>
</citation>
</ref>
<ref id="B33">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Mitchell</surname> <given-names>P. S.</given-names>
</name>
<name>
<surname>Roncaioli</surname> <given-names>J. L.</given-names>
</name>
<name>
<surname>Turcotte</surname> <given-names>E. A.</given-names>
</name>
<name>
<surname>Goers</surname> <given-names>L.</given-names>
</name>
<name>
<surname>Chavez</surname> <given-names>R. A.</given-names>
</name>
<name>
<surname>Lee</surname> <given-names>A. Y.</given-names>
</name>
<etal/>
</person-group>. (<year>2020</year>). <article-title>NAIP-NLRC4-deficient mice are susceptible to shigellosis</article-title>. <source>Elife</source> <volume>9</volume>, <elocation-id>e59022</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.7554/eLife.59022</pub-id>
</citation>
</ref>
<ref id="B34">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Morioka</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Nigorikawa</surname> <given-names>K.</given-names>
</name>
<name>
<surname>Okada</surname> <given-names>E.</given-names>
</name>
<name>
<surname>Tanaka</surname> <given-names>Y.</given-names>
</name>
<name>
<surname>Kasuu</surname> <given-names>Y.</given-names>
</name>
<name>
<surname>Yamada</surname> <given-names>M.</given-names>
</name>
<etal/>
</person-group>. (<year>2018</year>). <article-title>TMEM55a localizes to macrophage phagosomes to downregulate phagocytosis</article-title>. <source>J. Cell Sci.</source> <volume>131</volume>, <fpage>jcs213272</fpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1242/jcs.213272</pub-id>
</citation>
</ref>
<ref id="B35">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Mounier</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Laurent</surname> <given-names>V.</given-names>
</name>
<name>
<surname>Hall</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Fort</surname> <given-names>P.</given-names>
</name>
<name>
<surname>Carlier</surname> <given-names>M. F.</given-names>
</name>
<name>
<surname>Sansonetti</surname> <given-names>P. J.</given-names>
</name>
<etal/>
</person-group>. (<year>1999</year>). <article-title>Rho family GTPases control entry of shigella flexneri into epithelial cells but not intracellular motility</article-title>. <source>J. Cell Sci.</source> <volume>112</volume> (<issue>Pt 13</issue>), <fpage>2069</fpage>&#x2013;<lpage>2080</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1242/jcs.112.13.2069</pub-id>
</citation>
</ref>
<ref id="B36">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Muthuramalingam</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Whittier</surname> <given-names>S. K.</given-names>
</name>
<name>
<surname>Picking</surname> <given-names>W. L.</given-names>
</name>
<name>
<surname>Picking</surname> <given-names>W. D.</given-names>
</name>
</person-group> (<year>2021</year>). <article-title>The shigella type III secretion system: An overview from top to bottom</article-title>. <source>Microorganisms</source> <volume>9</volume>, <elocation-id>451</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.3390/microorganisms9020451</pub-id>
</citation>
</ref>
<ref id="B37">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Niebuhr</surname> <given-names>K.</given-names>
</name>
<name>
<surname>Giuriato</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Pedron</surname> <given-names>T.</given-names>
</name>
<name>
<surname>Philpott</surname> <given-names>D. J.</given-names>
</name>
<name>
<surname>Gaits</surname> <given-names>F.</given-names>
</name>
<name>
<surname>Sable</surname> <given-names>J.</given-names>
</name>
<etal/>
</person-group>. (<year>2002</year>). <article-title>Conversion of PtdIns(4,5)P(2) into PtdIns(5)P by the s.flexneri effector IpgD reorganizes host cell morphology</article-title>. <source>EMBO J.</source> <volume>21</volume>, <fpage>5069</fpage>&#x2013;<lpage>5078</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1093/emboj/cdf522</pub-id>
</citation>
</ref>
<ref id="B38">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Norris</surname> <given-names>F. A.</given-names>
</name>
<name>
<surname>Wilson</surname> <given-names>M. P.</given-names>
</name>
<name>
<surname>Wallis</surname> <given-names>T. S.</given-names>
</name>
<name>
<surname>Galyov</surname> <given-names>E. E.</given-names>
</name>
<name>
<surname>Majerus</surname> <given-names>P. W.</given-names>
</name>
</person-group> (<year>1998</year>). <article-title>SopB, a protein required for virulence of salmonella dublin, is an inositol phosphate phosphatase</article-title>. <source>Proc. Natl. Acad. Sci. U.S.A.</source> <volume>95</volume>, <fpage>14057</fpage>&#x2013;<lpage>14059</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1073/pnas.95.24.14057</pub-id>
</citation>
</ref>
<ref id="B39">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Patel</surname> <given-names>J. C.</given-names>
</name>
<name>
<surname>Hueffer</surname> <given-names>K.</given-names>
</name>
<name>
<surname>Lam</surname> <given-names>T. T.</given-names>
</name>
<name>
<surname>Gal&#xe1;n</surname> <given-names>J. E.</given-names>
</name>
</person-group> (<year>2009</year>). <article-title>Diversification of a salmonella virulence protein function by ubiquitin-dependent differential localization</article-title>. <source>Cell</source> <volume>137</volume>, <fpage>283</fpage>&#x2013;<lpage>294</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.cell.2009.01.056</pub-id>
</citation>
</ref>
<ref id="B40">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Payrastre</surname> <given-names>B.</given-names>
</name>
<name>
<surname>Gaits-Iacovoni</surname> <given-names>F.</given-names>
</name>
<name>
<surname>Sansonetti</surname> <given-names>P.</given-names>
</name>
<name>
<surname>Tronch&#xe8;re</surname> <given-names>H.</given-names>
</name>
</person-group> (<year>2012</year>). <article-title>Phosphoinositides and cellular pathogens</article-title>. <source>Subcell Biochem.</source> <volume>59</volume>, <fpage>363</fpage>&#x2013;<lpage>388</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1007/978-94-007-3015-1_12</pub-id>
</citation>
</ref>
<ref id="B41">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Pendaries</surname> <given-names>C.</given-names>
</name>
<name>
<surname>Tronch&#xe8;re</surname> <given-names>H.</given-names>
</name>
<name>
<surname>Arbibe</surname> <given-names>L.</given-names>
</name>
<name>
<surname>Mounier</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Gozani</surname> <given-names>O.</given-names>
</name>
<name>
<surname>Cantley</surname> <given-names>L.</given-names>
</name>
<etal/>
</person-group>. (<year>2006</year>). <article-title>PtdIns5P activates the host cell PI3-kinase/Akt pathway during shigella flexneri infection</article-title>. <source>EMBO J.</source> <volume>25</volume>, <fpage>1024</fpage>&#x2013;<lpage>1034</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/sj.emboj.7601001</pub-id>
</citation>
</ref>
<ref id="B42">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Piscatelli</surname> <given-names>H. L.</given-names>
</name>
<name>
<surname>Li</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Zhou</surname> <given-names>D.</given-names>
</name>
</person-group> (<year>2016</year>). <article-title>Dual 4- and 5-phosphatase activities regulate SopB-dependent phosphoinositide dynamics to promote bacterial entry</article-title>. <source>Cell Microbiol.</source> <volume>18</volume>, <fpage>705</fpage>&#x2013;<lpage>719</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1111/cmi.12542</pub-id>
</citation>
</ref>
<ref id="B43">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Pizarro-Cerd&#xe1;</surname> <given-names>J.</given-names>
</name>
<name>
<surname>K&#xfc;hbacher</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Cossart</surname> <given-names>P.</given-names>
</name>
</person-group> (<year>2015</year>). <article-title>Phosphoinositides and host-pathogen interactions</article-title>. <source>Biochim. Biophys. Acta</source> <volume>1851</volume>, <fpage>911</fpage>&#x2013;<lpage>918</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.bbalip.2014.09.011</pub-id>
</citation>
</ref>
<ref id="B44">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Popa</surname> <given-names>C. M.</given-names>
</name>
<name>
<surname>Tabuchi</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Valls</surname> <given-names>M.</given-names>
</name>
</person-group> (<year>2016</year>). <article-title>Modification of bacterial effector proteins inside eukaryotic host cells</article-title>. <source>Front. Cell Infect. Microbiol.</source> <volume>6</volume>. doi:&#xa0;<pub-id pub-id-type="doi">10.3389/fcimb.2016.00073</pub-id>
</citation>
</ref>
<ref id="B45">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Puhar</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Tronch&#xe8;re</surname> <given-names>H.</given-names>
</name>
<name>
<surname>Payrastre</surname> <given-names>B.</given-names>
</name>
<name>
<surname>Nhieu</surname> <given-names>G. T. V.</given-names>
</name>
<name>
<surname>Sansonetti</surname> <given-names>P. J.</given-names>
</name>
</person-group> (<year>2013</year>). <article-title>A shigella effector dampens inflammation by regulating epithelial release of danger signal ATP through production of the lipid mediator PtdIns5P</article-title>. <source>Immunity</source> <volume>39</volume>, <fpage>1121</fpage>&#x2013;<lpage>1131</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.immuni.2013.11.013</pub-id>
</citation>
</ref>
<ref id="B46">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Ramel</surname> <given-names>D.</given-names>
</name>
<name>
<surname>Lagarrigue</surname> <given-names>F.</given-names>
</name>
<name>
<surname>Pons</surname> <given-names>V.</given-names>
</name>
<name>
<surname>Mounier</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Dupuis-Coronas</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Chicanne</surname> <given-names>G.</given-names>
</name>
<etal/>
</person-group>. (<year>2011</year>). <article-title>Shigella flexneri infection generates the lipid PI5P to alter endocytosis and prevent termination of EGFR signaling</article-title>. <source>Sci. Signal</source> <volume>4</volume>, <fpage>ra61</fpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1126/scisignal.2001619</pub-id>
</citation>
</ref>
<ref id="B47">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Rey</surname> <given-names>C.</given-names>
</name>
<name>
<surname>Chang</surname> <given-names>Y.-Y.</given-names>
</name>
<name>
<surname>Latour-Lambert</surname> <given-names>P.</given-names>
</name>
<name>
<surname>Varet</surname> <given-names>H.</given-names>
</name>
<name>
<surname>Proux</surname> <given-names>C.</given-names>
</name>
<name>
<surname>Legendre</surname> <given-names>R.</given-names>
</name>
<etal/>
</person-group>. (<year>2020</year>). <article-title>Transcytosis subversion by m cell-to-enterocyte spread promotes shigella flexneri and listeria monocytogenes intracellular bacterial dissemination</article-title>. <source>PloS Pathog.</source> <volume>16</volume>, <elocation-id>e1008446</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.1371/journal.ppat.1008446</pub-id>
</citation>
</ref>
<ref id="B48">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Scanu</surname> <given-names>T.</given-names>
</name>
<name>
<surname>Spaapen</surname> <given-names>R. M.</given-names>
</name>
<name>
<surname>Bakker</surname> <given-names>J. M.</given-names>
</name>
<name>
<surname>Pratap</surname> <given-names>C. B.</given-names>
</name>
<name>
<surname>Wu</surname> <given-names>L.</given-names>
</name>
<name>
<surname>Hofland</surname> <given-names>I.</given-names>
</name>
<etal/>
</person-group>. (<year>2015</year>). <article-title>Salmonella manipulation of host signaling pathways provokes cellular transformation associated with gallbladder carcinoma</article-title>. <source>Cell Host Microbe</source> <volume>17</volume>, <fpage>763</fpage>&#x2013;<lpage>774</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.chom.2015.05.002</pub-id>
</citation>
</ref>
<ref id="B49">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Schr&#xf6;dinger</surname> <given-names>L.</given-names>
</name>
<name>
<surname>DeLano</surname> <given-names>W.</given-names>
</name>
</person-group> (<year>2020</year>) <source>PyMOL</source>, Available at: <uri xlink:href="http://www.pymol.org/pymol">http://www.pymol.org/pymol</uri>.</citation>
</ref>
<ref id="B50">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Steele-Mortimer</surname> <given-names>O.</given-names>
</name>
<name>
<surname>Knodler</surname> <given-names>L. A.</given-names>
</name>
<name>
<surname>Marcus</surname> <given-names>S. L.</given-names>
</name>
<name>
<surname>Scheid</surname> <given-names>M. P.</given-names>
</name>
<name>
<surname>Goh</surname> <given-names>B.</given-names>
</name>
<name>
<surname>Pfeifer</surname> <given-names>C. G.</given-names>
</name>
<etal/>
</person-group>. (<year>2000</year>). <article-title>Activation of akt/protein kinase b in epithelial cells by the salmonella typhimurium effector sigD</article-title>. <source>J. Biol. Chem.</source> <volume>275</volume>, <fpage>37718</fpage>&#x2013;<lpage>37724</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1074/jbc.M008187200</pub-id>
</citation>
</ref>
<ref id="B51">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>St&#xe9;venin</surname> <given-names>V.</given-names>
</name>
<name>
<surname>Chang</surname> <given-names>Y.-Y.</given-names>
</name>
<name>
<surname>Le Toquin</surname> <given-names>Y.</given-names>
</name>
<name>
<surname>Duchateau</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Gianetto</surname> <given-names>Q. G.</given-names>
</name>
<name>
<surname>Luk</surname> <given-names>C. H.</given-names>
</name>
<etal/>
</person-group>. (<year>2019</year>). <article-title>Dynamic growth and shrinkage of the salmonella-containing vacuole determines the intracellular pathogen niche</article-title>. <source>Cell Rep.</source> <volume>29</volume>, <fpage>3958</fpage>&#x2013;<lpage>3973.e7</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.celrep.2019.11.049</pub-id>
</citation>
</ref>
<ref id="B52">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>St&#xe9;venin</surname> <given-names>V.</given-names>
</name>
<name>
<surname>Giai Gianetto</surname> <given-names>Q.</given-names>
</name>
<name>
<surname>Duchateau</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Matondo</surname> <given-names>M.</given-names>
</name>
<name>
<surname>Enninga</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Chang</surname> <given-names>Y.-Y.</given-names>
</name>
</person-group> (<year>2021</year>). <article-title>Purification of infection-associated macropinosomes by magnetic isolation for proteomic characterization</article-title>. <source>Nat. Protoc.</source> <volume>16</volume>, <fpage>5220</fpage>&#x2013;<lpage>5249</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/s41596-021-00610-5</pub-id>
</citation>
</ref>
<ref id="B53">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Sun</surname> <given-names>C. H.</given-names>
</name>
<name>
<surname>Wacquier</surname> <given-names>B.</given-names>
</name>
<name>
<surname>Aguilar</surname> <given-names>D. I.</given-names>
</name>
<name>
<surname>Carayol</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Denis</surname> <given-names>K.</given-names>
</name>
<name>
<surname>Boucherie</surname> <given-names>S.</given-names>
</name>
<etal/>
</person-group>. (<year>2017</year>). <article-title>The shigella type III effector IpgD recodes Ca2+ signals during invasion of epithelial cells</article-title>. <source>EMBO J.</source> <volume>36</volume>, <fpage>2567</fpage>&#x2013;<lpage>2580</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.15252/embj.201696272</pub-id>
</citation>
</ref>
<ref id="B54">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Tahoun</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Mahajan</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Paxton</surname> <given-names>E.</given-names>
</name>
<name>
<surname>Malterer</surname> <given-names>G.</given-names>
</name>
<name>
<surname>Donaldson</surname> <given-names>D. S.</given-names>
</name>
<name>
<surname>Wang</surname> <given-names>D.</given-names>
</name>
<etal/>
</person-group>. (<year>2012</year>). <article-title>Salmonella transforms follicle-associated epithelial cells into m cells to promote intestinal invasion</article-title>. <source>Cell Host Microbe</source> <volume>12</volume>, <fpage>645</fpage>&#x2013;<lpage>656</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.chom.2012.10.009</pub-id>
</citation>
</ref>
<ref id="B55">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Tran Van Nhieu</surname> <given-names>G.</given-names>
</name>
<name>
<surname>Kai Liu</surname> <given-names>B.</given-names>
</name>
<name>
<surname>Zhang</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Pierre</surname> <given-names>F.</given-names>
</name>
<name>
<surname>Prigent</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Sansonetti</surname> <given-names>P.</given-names>
</name>
<etal/>
</person-group>. (<year>2013</year>). <article-title>Actin-based confinement of calcium responses during shigella invasion</article-title>. <source>Nat. Commun.</source> <volume>4</volume> (<issue>1567</issue>), <page-range>1&#x2013;10</page-range>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/ncomms2561</pub-id>
</citation>
</ref>
<ref id="B56">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Valencia-Gallardo</surname> <given-names>C. M.</given-names>
</name>
<name>
<surname>Carayol</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Tran Van Nhieu</surname> <given-names>G.</given-names>
</name>
</person-group> (<year>2015</year>). <article-title>Cytoskeletal mechanics during shigella invasion and dissemination in epithelial cells</article-title>. <source>Cell Microbiol.</source> <volume>17</volume>, <fpage>174</fpage>&#x2013;<lpage>182</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1111/cmi.12400</pub-id>
</citation>
</ref>
<ref id="B57">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Viaud</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Lagarrigue</surname> <given-names>F.</given-names>
</name>
<name>
<surname>Ramel</surname> <given-names>D.</given-names>
</name>
<name>
<surname>Allart</surname> <given-names>S.</given-names>
</name>
<name>
<surname>Chicanne</surname> <given-names>G.</given-names>
</name>
<name>
<surname>Ceccato</surname> <given-names>L.</given-names>
</name>
<etal/>
</person-group>. (<year>2014</year>). <article-title>Phosphatidylinositol 5-phosphate regulates invasion through binding and activation of Tiam1</article-title>. <source>Nat. Commun.</source> <volume>5</volume> (<issue>4080</issue>), <page-range>1&#x2013;17</page-range>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/ncomms5080</pub-id>
</citation>
</ref>
<ref id="B58">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Walpole</surname> <given-names>G. F. W.</given-names>
</name>
<name>
<surname>Pacheco</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Chauhan</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Clark</surname> <given-names>J.</given-names>
</name>
<name>
<surname>Anderson</surname> <given-names>K. E.</given-names>
</name>
<name>
<surname>Abbas</surname> <given-names>Y. M.</given-names>
</name>
<etal/>
</person-group>. (<year>2022</year>). <article-title>Kinase-independent synthesis of 3-phosphorylated phosphoinositides by a phosphotransferase</article-title>. <source>Nat. Cell Biol.</source> <volume>24</volume>, <fpage>708</fpage>&#x2013;<lpage>722</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/s41556-022-00895-y</pub-id>
</citation>
</ref>
<ref id="B59">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Wang</surname> <given-names>L.</given-names>
</name>
<name>
<surname>Zhong</surname> <given-names>H.</given-names>
</name>
<name>
<surname>Xue</surname> <given-names>Z.</given-names>
</name>
<name>
<surname>Wang</surname> <given-names>Y.</given-names>
</name>
</person-group> (<year>2022</year>). <article-title>Improving the topology prediction of &#x3b1;-helical transmembrane proteins with deep transfer learning</article-title>. <source>Comput. Struct. Biotechnol. J.</source> <volume>20</volume>, <fpage>1993</fpage>&#x2013;<lpage>2000</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.csbj.2022.04.024</pub-id>
</citation>
</ref>
<ref id="B60">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Weddle</surname> <given-names>E.</given-names>
</name>
<name>
<surname>Agaisse</surname> <given-names>H.</given-names>
</name>
</person-group> (<year>2018</year>). <article-title>Principles of intracellular bacterial pathogen spread from cell to cell</article-title>. <source>PloS Pathog.</source> <volume>14</volume>, <fpage>e1007380</fpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1371/journal.ppat.1007380</pub-id>
</citation>
</ref>
<ref id="B61">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Weigele</surname> <given-names>B. A.</given-names>
</name>
<name>
<surname>Orchard</surname> <given-names>R. C.</given-names>
</name>
<name>
<surname>Jimenez</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Cox</surname> <given-names>G. W.</given-names>
</name>
<name>
<surname>Alto</surname> <given-names>N. M.</given-names>
</name>
</person-group> (<year>2017</year>). <article-title>A systematic exploration of the interactions between bacterial effector proteins and host cell membranes</article-title>. <source>Nat. Commun.</source> <volume>8</volume>, <fpage>532</fpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1038/s41467-017-00700-7</pub-id>
</citation>
</ref>
<ref id="B62">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Weiner</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Enninga</surname> <given-names>J.</given-names>
</name>
</person-group> (<year>2019</year>). <article-title>The pathogen-host interface in three dimensions: Correlative FIB/SEM applications</article-title>. <source>Trends Microbiol.</source> <volume>27</volume>, <fpage>426</fpage>&#x2013;<lpage>439</lpage>. doi:&#xa0;<pub-id pub-id-type="doi">10.1016/j.tim.2018.11.011</pub-id>
</citation>
</ref>
<ref id="B63">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Weiner</surname> <given-names>A.</given-names>
</name>
<name>
<surname>Mellouk</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Lopez-Montero</surname> <given-names>N.</given-names>
</name>
<name>
<surname>Chang</surname> <given-names>Y.-Y.</given-names>
</name>
<name>
<surname>Souque</surname> <given-names>C.</given-names>
</name>
<name>
<surname>Schmitt</surname> <given-names>C.</given-names>
</name>
<etal/>
</person-group>. (<year>2016</year>). <article-title>Macropinosomes are key players in early shigella invasion and vacuolar escape in epithelial cells</article-title>. <source>PloS Pathog.</source> <volume>12</volume>, <elocation-id>e1005602</elocation-id>. doi:&#xa0;<pub-id pub-id-type="doi">10.1371/journal.ppat.1005602</pub-id>
</citation>
</ref>
</ref-list>
</back>
</article>