Original Research ARTICLE
The conserved ESCRT III machinery participates in the phagocytosis of Entamoeba histolytica
- 1Theory and Bio-Systems, Max Planck Institute of Colloids and Interfaces (MPG), Germany
- 2Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico
The endosomal sorting complex required for transport (ESCRT) orchestrates cell membrane-remodeling mechanisms in eukaryotes, including endocytosis. However, ESCRT functions in phagocytosis(ingestion of ≥ 250 nm particles), has been poorly studied. In macrophages and amoebae, phagocytosis is required for cell nutrition and attack to other microorganisms and cells.In Entamoeba histolytica, the voracious protozoan responsible for human amoebiasis, phagocytosisis a land mark of virulence. Here, we have investigated the role of ESCRT-III in the phagocytosis of E. histolytica, using mutant trophozoites, recombinant proteins (rEhVps20, rEhVps32, rEhVps24 and rEhVps2) and giant unilamellar vesicles (GUVs). Confocal images displayed the four proteins located around the ingested erythrocytes,in erythrocytes-containing phagosomes and in multivesicular bodies. EhVps32 and EhVps2 proteins co-localized at the phagocytic cups.Protein association increased during phagocytosis. Immunoprecipitation and flow cytometry assays substantiated these associations. GUVs revealed that the protein assembly sequence is essential to form intraluminal vesicles (ILVs). First, the active rEhVps20 bound to membranes and recruited rEhVps32, promoting membrane invaginations. rEhVps24 allowed the detachment of nascent vesicles, forming ILVs; and rEhVps2 modulated their size. The knock down of Ehvps20 and Ehvps24genes diminished the rate of erythrophagocytosis demonstrating the importance of ESCRT-III in this event. In conclusion, we present here evidence of the ESCRT-III participation in phagocytosis and delimitate the putative function of proteins, according to thein vitro reconstruction of their assembling.
Keywords: Protozoan parasites, ESCRT-III proteins, Entamoeba histolytica, Phagocytosis, GUVs model
Received: 11 Dec 2017;
Accepted: 12 Feb 2018.
Edited by:Anjan Debnath, University of California, San Diego, United States
Reviewed by:César López-Camarillo, Universidad Autónoma de la Ciudad de México, Mexico
Lesly Temesvari, Clemson University, United States
Copyright: © 2018 Avalos-Padilla, Knorr, Javier-Reyna, García-Rivera, Lipowsky, Dimova and Orozco. 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 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.
* Correspondence: Dr. Esther Orozco, PROFESSOR., Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Infectómica y Patogénesis Molecular, Ave IPN #2508, Mexico City, Ave IPN #2508, México, 0736', Ciudad de México, Mexico, firstname.lastname@example.org