Abstract
Zika virus (ZIKV) causes severe neurological disease, including microcephaly and Guillain-Barré syndrome, through complex interactions with host cell proteins. This review synthesizes the 2015–2025 published literature on ZIKV-host protein interactions and their therapeutic targeting. ZIKV enters cells via multiple receptor pathways: adhesion receptors (DC-SIGN, Hsp70), high-affinity entry receptors (ITGB4, GRP78, NCAM1), internalization receptors (integrin αvβ5, sialic acid), and endosomal receptors (AXL, TIM-1, CD300a). Viral structural proteins direct virion assembly, while nonstructural proteins NS1–NS5 suppress immune responses, remodel cellular membranes, and dysregulate gene expression. NS5 uniquely suppresses neurodevelopmental genes and disrupts ciliary function through nuclear localization, directly driving microcephaly pathogenesis. Therapeutic strategies include receptor antagonists, protease inhibitors, and polymerase inhibitors. However, receptor redundancy, viral protein multifunctionality, and pregnancy safety constraints limit clinical translation. This review identifies ZIKV-host protein interactions as therapeutic targets and highlights barriers to drug development.
1 Introduction
Zika virus (ZIKV) is a mosquito-borne flavivirus first isolated in 1947 from a rhesus monkey in the Zika Forest, Uganda. For nearly six decades, ZIKV caused only sporadic infections with mild symptoms. The 2015–2016 outbreak in the Americas fundamentally altered this epidemiological pattern (). ZIKV rapidly spread from Brazil throughout Central and South America, infecting millions and causing severe neurological complications. The outbreak revealed a devastating association between maternal ZIKV infection and congenital Zika syndrome, which is characterized by severe microcephaly, brain malformations, and neurodevelopmental abnormalities in newborns (). ZIKV infection also caused Guillain-Barré syndrome and other neurological complications in adults (). In 2016, the World Health Organization declared the outbreak a Public Health Emergency of International Concern.
Zika virus causes disease by utilizing different host cell receptors for cellular entry and by establishing intricate interactions with host proteins following entry (). These protein interactions enable ZIKV to infect diverse cell types, including those in the fetal brain and placenta, and to manipulate host pathways that support viral replication while suppressing immune responses. This capacity to exploit multiple entry routes and commandeer cellular machinery is central to ZIKV dissemination throughout the body and the ensuing neurological and developmental pathology. Understanding these interactions is critical for identifying novel therapeutic strategies and for limiting the severity and scope of ZIKV infection.
Targeting ZIKV-host protein interactions represents a highly promising antiviral strategy that offers considerable advantages over traditional direct-acting antivirals (). This approach reduces the likelihood of drug resistance because host proteins are genetically stable and do not mutate like viral genomes, and it provides broad-spectrum activity since many host factors are exploited by multiple flaviviruses. ZIKV depends extensively on host cellular machinery throughout its life cycle, creating numerous therapeutic intervention points from viral entry through replication and assembly. Entry can be blocked by targeting multiple entry receptors. Structural protein interactions can be disrupted to impair capsid function and block virion assembly. Nonstructural proteins offer additional targets (; ).
2 ZIKV receptors
ZIKV infection proceeds through multiple, sequential stages involving diverse host receptors that perform distinct functional roles at different infection steps. This classification organizes confirmed and candidate ZIKV receptors according to their mechanistic contributions during viral entry and infection, from initial adhesion through membrane fusion (Figure 1).
Figure 1
2.1 Adhesion receptors
Adhesion receptors mediate initial, low-affinity interactions between viral particles and the cell surface, representing the first step of viral entry. Dysfunction of these receptors typically delays but does not eliminate infection, distinguishing them functionally from high-affinity entry receptors.
Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin (DC-SIGN) is a C-type lectin receptor highly expressed on immature macrophages and dendritic cells. It serves as an attachment factor for multiple flaviviruses including DENV, WNV, JEV, and ZIKV, all of which display mannose-rich glycans recognized by the DC-SIGN (). The N-terminus carbohydrate recognition domain (CRD) of DC-SIGN recognizes N-linked glycosylation sites on the ZIKV envelope protein in a calcium-dependent and mannose/fucose-selective manner (). This binding is specifically inhibited by anti-DC-SIGN antibodies, mannose competitors, or calcium chelators (EDTA).
Heat shock protein 70 (Hsp70) functions as a cell surface attachment factor facilitating ZIKV entry (). Hsp70 translocates to the plasma membrane where it directly interacts with the ZIKV envelope protein. Hsp70 helps attach and concentrate ZIKV particles at the cell surface, and increase engagement with high-affinity entry receptors (). Anti-Hsp70 antibodies significantly reduce ZIKV infection rates and plaque-forming units. Co-incubation of ZIKV with recombinant Hsp70 protein decreases ZIKV particle copy numbers in culture supernatants. Magnetic nanosensor studies further confirm direct binding interactions between Hsp70 and the ZIKV envelope protein (). Mechanistically, Hsp70 likely facilitates clathrin-mediated endocytosis, as Hsp70 localizes to clathrin-coated pits at the plasma membrane.
Glycosaminoglycans (GAGs) exhibit a paradoxical, context-dependent role in ZIKV infection that distinguishes it from related flaviviruses. Although ZIKV envelope protein binding pharmaceutical heparin with nanomolar affinity (KD = 443 nM for various sulfated GAGs) through electrostatically-driven, chain-length-dependent interactions with sulfated GAG motifs (). However, multiple independent genome-wide CRISPR/Cas9 knockout screens definitively demonstrate that heparan sulfate (HS) biosynthesis gene deficiency (including EXT1, EXT2, EXTL3) does not significantly impair ZIKV attachment, internalization, or overall viral replication (). This distinguishes ZIKV fundamentally from related flaviviruses such as DENV and JEV, which authentically depend on HS as obligate attachment co-receptors. It was proposed that HS promotes early viral replication by maintaining autophagy that ZIKV hijacks to establish replication organelles, but later restricts infection by targeting viral envelope and NS3 proteins for proteasome-dependent degradation ().
2.2 High-affinity entry receptors
Following initial adhesion, entry receptors establish high-affinity binding between virus and cell surface, directly mediating productive viral entry. These receptors are the primary determinants of cellular tropism and cell-type-specific infection.
Integrin β4 (ITGB4, CD104) forms heterodimers with integrin α6 (ITGA6) on cell surfaces and serves as a critical entry receptor for ZIKV. ITGB4 demonstrates virus-specific selectivity. It mediates ZIKV entry but not JEV entry, distinguishing ZIKV from other flaviviruses that preferentially utilize integrin αvβ3. Soluble recombinant ITGB4 proteins and anti-ITGB4 antibody significantly reduced ZIKV binding and infection in a dose-dependent manner. In vivo studies further demonstrated that anti-ITGB4 antibodies prevented ZIKV infection in placental tissue, reduced viral titers in embryos, and increased embryo survival rates in pregnant mice ().
Glucose-Regulated Protein 78 (GRP78) functions as a critical cell surface receptor facilitating ZIKV entry through direct binding to the ZIKV envelope protein domain III (EDIII) (Wang S. et al., 2020). GRP78 translocated from the ER lumen to the cell surface, where it serves as an attachment factor. The interaction involves both the nucleotide-binding domain (NBD) and substrate-binding domain (SBD) of GRP78, with critical residues (R492, T518) in the SBD mediating a canonical chaperone-client binding mechanism. Functional blocking experiments reveal that monoclonal antibodies targeting the N-terminus of GRP78 inhibit ZIKV entry by 50–70%, while siRNA-mediated GRP78 knockdown produces similar reductions. Following GRP78-mediated attachment, ZIKV enters cells predominantly through clathrin-mediated endocytosis, trafficking through Rab5+ early endosomes to Rab7+ late endosomes where low pH triggers E protein conformational changes and subsequent membrane fusion ().
Neural Cell Adhesion Molecule 1 (NCAM1/CD56) is an immunoglobulin superfamily glycoprotein highly expressed in brain tissue, neurons, glia, and skeletal muscle that functions as a high-affinity ZIKV entry receptor identified through time-resolved chemical proteomics (). Multiple lines of evidence support this role, including direct binding of NCAM1 to the ZIKV envelope protein demonstrated via co-immunoprecipitation, blocking of ZIKV infection by the recombinant NCAM1 extracellular domain, reduced ZIKV infection upon anti-NCAM1 antibody treatment, enhanced ZIKV infection in NCAM1-overexpressing HEK 293T cells, and marked attenuation of ZIKV infection in CRISPR/Cas9-mediated NCAM1 knockout U-251 MG glioblastoma cells.
2.3 Internalization receptors
Internalization receptors specifically mediate viral internalization after virus-cell binding is established, rather than facilitating initial attachment. Integrin αvβ5 was identified through genome-wide CRISPR-Cas9 screening as a specialized receptor mediating ZIKV infection in neural stem cells (Zhu et al., 2020). Integrin αvβ5 specifically mediates ZIKV internalization rather than initial attachment (Wang S. et al., 2020). αvβ5 directly binds ZIKV virions and activates focal adhesion kinase (FAK), which is essential for productive viral infection. The expression level of αvβ5 correlates with ZIKV susceptibility in neural tissues and exhibits specific tropism patterns in the developing human cerebral cortex, with particularly high expression in neural stem cells. Notably, αvβ5-blocking antibodies and small-molecule inhibitors such as SB273005 and cilengitide effectively reduce ZIKV infection and alleviate virus-induced pathology in human neural stem cells and mouse brain tissue ().
Cell surface α2,3-linked sialic acid facilitates ZIKV internalization rather than attachment (Zhu et al., 2020). Neuraminidase treatment removing sialic acid significantly reduces ZIKV infection in Vero cells, neural progenitor cells, and Huh7 cells. Sialic acid is dispensable for viral attachment at 4°C during inactive endocytosis but essential for internalization at 37°C, demonstrating its role in endocytic uptake rather than binding. ZIKV does not directly interact with sialic acid, as sialyllactose fails to inhibit infection. Instead, sialic acid facilitates endocytosis-mediated entry. ZIKV specifically requires α2,3-linked rather than α2,6-linked sialic acid, as confirmed by linkage-specific neuraminidase treatments and ST3GAL4 knockout studies.
2.4 Endosomal receptors
Endosomal receptors localize to endosomal membranes and function under acidic pH conditions to mediate membrane fusion processes. AXL, a TAM receptor family member (TYRO3, AXL, MERTK), plays a complex and controversial role in ZIKV entry. Expression profiling showed positive correlation between AXL expression and ZIKV tropism in neural cells (). As a candidate receptor, AXL indirectly binds ZIKV via Gas6 and mediates viral entry through clathrin-mediated endocytosis to Rab5+ endosomes. The ZIKV/Gas6/AXL complex suppresses interferon signaling by activating AXL kinase, inhibiting ISG expression and type I interferon production, thereby facilitating ZIKV replication (; Zwernik et al., 2021). However, AXL’s role as an essential ZIKV receptor remains controversial, while AXL knockout neural precursor cells remained susceptible to ZIKV infection (Wells et al., 2016). Pregnant mouse models revealed comparable ZIKV RNA levels across wild-type, AXL knockout, and MERTK knockout mice, indicating that AXL and MERTK are not essential for ZIKV infection in IFNAR-blocked mice (). Furthermore, a single PrM gene mutation (H83R) determines viral AXL-dependence, enabling AXL-independent infection of human neural cells and suggesting that AXL contributions vary by viral strain and cell type ().
Phosphatidylserine (PS) receptors play critical roles in late-stage ZIKV infection. T-cell immunoglobulin and mucin domain-containing protein 1 (TIM-1) has been confirmed as an important PS receptor enhancing ZIKV entry into cells. Cell electrical impedance biosensing studies demonstrate that ZIKV entry is highly sensitive to subtle changes in TIM-1 expression levels: both overexpression of TIM-1 in infection-resistant HEK293T cells and partial knockout of TIM-1 in susceptible A549 cells significantly modulate viral infection efficiency (Yu et al., 2025). Additionally, MARCH2 and MARCH3 function as redundant host restriction factors by targeting TIM-1 for K48-linked polyubiquitination and proteasomal degradation, thereby suppressing ZIKV infection (Zhang et al., 2025). CD300a represents another PS receptor recently confirmed to facilitate ZIKV cell entry (). Inhibition of CD300a in immature monocyte-derived dendritic cells partially but significantly reduces ZIKV replication. Collectively, these findings demonstrate that ZIKV exploits multiple PS receptors to achieve efficient cell entry.
2.5 Modulatory factors
ZIKV infection of blood-brain barrier endothelial cells upregulates VCAM-1, ICAM-1, and E-selectin at mRNA and protein levels, with soluble forms released 4–10 days post-infection that facilitate leukocyte recruitment and adhesion to the BBB (). Concurrently, ZIKV-infected primary human monocytes undergo proteomic reprogramming that upregulates integrins β1, α5, and αM; adhesion molecules ICAM-3, PECAM-1, CD99, and ITGAL; focal adhesion proteins including catenins, myosins, actinin, vinculin, talin, and filamins A/B; and the scaffolding protein IQGAP1 (). This adhesion molecule profile is ZIKV-specific, not replicated by HIV-1 infection or GM-CSF treatment, and functionally enhances monocyte attachment to endothelial cells and extracellular matrices. Coordinated upregulation of complementary adhesion molecules on both endothelial cells and infected monocytes, particularly CD14+ and CD16+ subsets, enables efficient monocyte transmigration across brain microvascular endothelial cells in transwell systems, with subsequent infection of basolateral astrocytes (). While ZIKV utilizes multiple receptor pathways for cell entry, comparative analysis across flaviviruses reveals both conserved and virus-specific receptor usage patterns (Table 1).
Table 1
| Receptor/Entry factor | Evidence quality | |||||
|---|---|---|---|---|---|---|
| ZIKV | DENV | WNV | JEV | YFV | TBEV | |
| ITGB4 | High | None | None | None | None | Not reported |
| ITGB3 | None | Low | High | High | Not reported | Not reported |
| ITGAV/ITGB5 | Medium | Not reported | Not reported | Not reported | Not reported | Not reported |
| ITGA6 with ITGB4 | Medium | Not reported | Not reported | Not reported | Not reported | Not reported |
| Heparan Sulfate | Low | High | High | High | Medium | Medium |
| Syndecan (HSPG variant) | Not studied | High | Medium | Low | Not reported | Medium |
| AXL (TAM family) | Medium | Low | Low | Low | Low | Not reported |
| MER (TAM family) | Low | Medium | Medium | Medium | Medium | Not reported |
| TYRO3 (TAM family) | Low | Medium | Low | Not reported | Not reported | Not reported |
| TIM-1 (PS receptor) | Medium | High | High | Not reported | Not reported | Not reported |
| TIM-4 (PS receptor) | Medium | High | High | Not reported | Not reported | Not reported |
| DC-SIGN (C-type lectin) | Medium | High | Medium | High | High | Not reported |
| SIGN (C-type lectin) | Medium | High | Low | High | High | Not reported |
| GRP78 (Hsp90 family) | Medium | High | Medium | High | Medium | Medium |
| Hsp70 (Hsp family) | Low | Medium | Medium | High | Medium | Medium |
| NCAM1 (CAM) | Medium | Not reported | Not reported | Not reported | Not reported | Not reported |
| Claudin-1 (tight junction) | Low | Medium | Not reported | Not reported | Not reported | Not reported |
| CD55 (Complement) | Low | Low | Medium | Not reported | High | Not reported |
| Phosphatidylserine (lipid) | High | High | High | Medium | Medium | Not reported |
| Clathrin (endocytosis) | High | High | High | High | High | High |
Comparative analysis of cellular receptor usage among flaviviruses.
3 Structural proteins interacting with host proteins
3.1 ZIKV capsid protein
ZIKV capsid protein orchestrates viral assembly through extensive interactions with host cellular machinery. Quantitative proteomics in mosquito cells identified 157 host protein interactors, with the transitional ER ATPase TER94 and its human ortholog valosin-containing protein (VCP) (). TER94/VCP, together with E3 ubiquitin ligase UBR5, targets capsid for ubiquitin-proteasome-dependent degradation in both mosquito and human cells, and this paradoxical turnover is required for efficient infection (). VCP inhibition markedly impairs viral replication and disrupts formation of replication organelles. Unlike dengue virus capsid, ZIKV capsid binds lipid droplets non-specifically but does not associate with very-low-density lipoproteins, linking capsid function to lipid-droplet-driven replication pathways ().
3.2 ZIKV prM protein
ZIKV precursor membrane protein prM engages multiple host factors to regulate viral replication, virion assembly, and pathogenicity. PIM1 kinase directly binds and phosphorylates prM at Ser101 and Thr107, preventing AMFR-mediated ubiquitination and proteasomal degradation, stabilizing prM and enhancing viral replication (). The prM transmembrane domain contains two functional cholesterol-binding motifs, where CARC2 supports cholesterol-dependent viral entry and CARC3 promotes virion assembly. Disruption of either motif severely impairs infection (). prM carries an essential N-linked glycosylation site required for efficient virion secretion and infectivity (). The ER chaperone GRP78/BiP facilitates prM folding, while furin protease cleaves prM in the trans-Golgi network after pH-induced conformational changes, generating both mature and partially mature virions.
3.3 ZIKV E protein
ZIKV E protein is a class II fusion glycoprotein with three ectodomains: DI acts as a central scaffold, DII harbors the fusion loop and undergoes conformational rearrangement during fusion, and DIII mediates receptor engagement and tropism (). A prominent N-linked glycan at Asn154 on the DI-DII hinge serves as an attachment site for DC-SIGN, which recognizes this glycan and mediates low-affinity initial adhesion (). ZIKV E binds highly sulfated glycosaminoglycans such as heparin with nanomolar, electrostatic and chain−length−dependent affinity, supporting a role for GAGs as adhesion factors on placental and neural cells (). Hsp70 translocates to the cell surface, binds ZIKV E protein, and functions as an attachment factor. GRP78 relocates to the cell surface, directly binds domain III of the ZIKV E ectodomain via its nucleotide−binding and substrate−binding domains, and promotes clathrin−mediated endocytosis and productive infection (). NCAM1 directly binds the ZIKV E ectodomain and acts as a high−affinity entry receptor in neural cells, with overexpression enhancing and knockout or blockade reducing viral binding and entry, thereby linking E–NCAM1 interaction to neural tropism and fetal brain infection ().
4 Non-structural proteins interacting with host proteins
ZIKV nonstructural proteins NS1–NS5 are generated through sequential proteolytic cleavage of the viral polyprotein and orchestrate diverse pathogenic mechanisms through extensive host protein interactions (Figure 2).
Figure 2
4.1 NS1 protein
ZIKV NS1 orchestrates a multifaceted host interaction network comprising at least 28 proteins organized into five functional modules. The immune evasion module includes USP8, which NS1 recruits to stabilize caspase-1, facilitating cGAS cleavage and suppressing type I interferon responses, while secreted NS1 interacts with complement components C1q, C4, C4BP, factor H, and vitronectin to antagonize complement-mediated virolysis (Zheng et al., 2018). The chaperone module consists of the cytosolic chaperonin TRiC-CCT complex and ER chaperone GRP78, which provide ATP-dependent folding and stabilization of NS1 at ER replication sites (Wang Y. et al., 2020). The organellar module comprises mitochondrial proteins and ER lipid-associated factors that NS1 uses to remodel ER membranes into replication factories and siphon healthy mitochondria from neighboring cells via tunneling nanotubes (). The cytoskeleton module includes actin, tubulin, and SUN2, which support tunneling nanotube formation and cytoskeletal reorganization required for NS1 trafficking and intercellular mitochondrial transfer. The stress response module involves RNase L and FMRP, RNA-binding proteins that NS1 co-opts to create a proviral environment (). Together, these interactions enable NS1 to suppress interferon responses, establish ER-derived replication factories, promote intercellular dissemination through tunneling nanotube-mediated transfer, and reprogram host stress and RNA metabolism pathways to support viral replication.
4.2 NS2A protein
ZIKV NS2A functions as a multifaceted virulence factor orchestrating host protein interactions across several interconnected networks. The NS2A-AMFR-FAM134B axis suppresses ER-phagy through K48-linked ubiquitination at lysine 56, targeting RETREG1/FAM134B for proteasomal degradation and maintaining ER membrane supply for viral replication organelles (Zhang et al., 2024). This mechanism is evidenced by attenuated microcephalic phenotypes in ZIKV-NS2A K56R mutants that cannot undergo ubiquitination. NS2A also mediates STAT1/STAT2 degradation via a proteasome-dependent pathway involving amino acids 12-100, preventing nuclear translocation despite active phosphorylation and effectively suppressing interferon-mediated antiviral responses (; ). In neural cells, NS2A disrupts adherens junction integrity through direct interactions with PARD3, CDH2/N-cadherin, CTNNB1/β-catenin, and CTNND1/p120-catenin, a mechanism uniquely absent in dengue virus NS2A that impairs neurogenesis and contributes to microcephaly in embryonic mouse cortex and human brain organoids (). NS2A also manipulates peroxisomal biogenesis via PEX3/PEX19 interactions to reprogram cellular lipid metabolism, as ZIKV infection depletes peroxisomes in human fetal astrocytes while peroxisome expansion restricts viral replication (Wong et al., 2019). Furthermore, NS2A RNA directly binds eIF2α to suppress proinflammatory cytokine translation, adding a post-transcriptional layer to immune evasion (Wu et al., 2024). Collectively, NS2A maintains ER homeostasis for replication, evades interferon and NF-κB signaling, disrupts neural adherens junctions and neurogenesis, and reprograms peroxisome and translational pathways, positioning NS2A as a central orchestrator of ZIKV neuropathogenesis.
4.3 NS2B-NS3 complex
The ZIKV NS2B-NS3 serine protease complex interacts with host proteins to drive neurotoxicity and immune evasion. In neural progenitor cells, NS2B-NS3 directly binds and cleaves the cytoskeletal GTPase Septin-2 at residue R306, causing cytokinesis failure, supernumerary centrosomes, prolonged mitosis, and cell death that contribute to microcephaly pathogenesis (). NS2B-NS3 also interacts with the ER oxidoreductase ERp57, which stabilizes the NS2B/NS3 complex, enhances viral replication, and promotes reactive oxygen species-mediated DNA damage and apoptosis. ERp57 depletion reduces ZIKV production and limits DNA damage, highlighting this host factor as a proviral cofactor (Wang et al., 2024). Additionally, NS2B-NS3 promotes proteasome-dependent degradation of JAK1, blocking type I interferon signaling and reducing interferon-stimulated gene induction. This effect is reinforced when NS2B-NS3 cooperates with NS1 and NS4B to prevent interferon-induced autophagic degradation of the protease complex (Wu et al., 2017b). Proteome-wide interaction mapping identifies additional NS2B-NS3 host partners involved in cytoskeleton organization, vesicle trafficking, and cell cycle control, establishing this protease complex as a central hub that couples viral polyprotein processing to host cell remodeling, antiviral pathway suppression, and neural cell injury (Tangsongcharoen et al., 2019; ).
4.4 NS4A
NS4A is an ER-resident protein that remodels intracellular membranes by recruiting host factors to form replication organelles and support viral RNA synthesis. NS4A directly interacts with the microcephaly protein ANKLE2 and co-opts its role in coordinating nuclear envelope and ER organization (). Loss of ANKLE2 or disruption of this complex impairs virus-induced membrane rearrangements and reduces ZIKV replication in mammalian and mosquito cells (). Genetic and functional studies show that NS4A-driven interference with ANKLE2-dependent pathways induces microcephaly-like phenotypes, linking this interaction to ZIKV-associated congenital brain malformations. Proteome-wide mapping indicates that NS4A also associates with ER membrane and stress-response factors involved in unfolded protein response signaling and ER homeostasis, consistent with the ER stress and UPR activation observed in ZIKV-infected neural progenitors (). These findings establish NS4A as a membrane-remodeling hub that hijacks ANKLE2 and other ER-associated proteins to build replication factories, evade cellular stress surveillance, and contribute to ZIKV neurotropism and microcephaly.
4.5 NS4B
NS4B interacts with host proteins to remodel membranes and suppress innate immunity, promoting efficient viral replication and persistence. NS4B binds the cholesterol biosynthesis enzyme DHCR7 and induces its expression, which inhibits TBK1 and IRF3 phosphorylation, reduces IFN-β and interferon-stimulated gene induction, and enhances ZIKV infection in human cells and mouse models (). Genetic or pharmacologic inhibition of DHCR7 restores antiviral signaling and restricts ZIKV replication. NS4B also blocks type I interferon induction by targeting the TBK1 signaling node, acting in parallel with NS1 and the NS2B-NS3 protease to dampen both interferon production and downstream JAK-STAT signaling (). Systematic mapping places NS4B within ER-resident protein networks involved in membrane protein biogenesis and stress responses, consistent with its role in shaping replication organelles and adjusting ER homeostasis during infection (; ). These interactions establish NS4B as a multifunctional antagonist of interferon pathways and a coordinator of membrane remodeling via DHCR7-dependent and ER-associated mechanisms.
4.6 NS5
NS5 is the largest and most conserved flavivirus protein, containing an N-terminal methyltransferase domain and a C-terminal RNA-dependent RNA polymerase domain that support viral RNA capping and replication while engaging over fifty host proteins in multiple functional networks (). Beyond its enzymatic roles, NS5 orchestrates immune evasion by promoting CRL3-ZSWIM8-dependent ubiquitination and proteasomal degradation of STAT2 to selectively block type I and type III interferon signaling while favoring type II interferon responses (). NS5 also uses its methyltransferase domain to bind RIG-I and inhibit K63-linked ubiquitination and interferon-β production, and cooperates with viral sfRNA to inhibit STAT1 phosphorylation and downstream signaling (; Wu et al., 2022). Uniquely for a cytoplasmically replicating RNA virus, NS5 accumulates in the nucleus where it binds chromatin at actively transcribed neural genes and suppresses their expression by interfering with PAF1 complex-mediated transcription elongation, providing a direct mechanism for ZIKV-induced impairment of neurodevelopment (; ). NS5 multimerizes at the base of primary and motile cilia to induce non-genetic ciliopathy characterized by shortened or lost cilia, premature neurogenesis, abnormal neuron delamination, and ependymal ciliary dysfunction. NS5 also relocalizes to centrosomes during mitosis to promote centrosome amplification, spindle defects, and mitotic abnormalities in neural progenitors. Additionally, NS5 directly binds the C-terminal region of p53 to activate p53-dependent transcription of pro-apoptotic genes and trigger apoptosis in human neural progenitor cells (). NS5 stability is regulated by the SUMO E3 ligase PIAS1, which stabilizes flavivirus NS5 through SUMOylation, and by the ubiquitin-activating enzyme UBA1, which promotes ZIKV replication and modulates NS5 ubiquitin dynamics (). NS5’s multifunctional activities spanning cytoplasmic replication, nuclear transcriptional repression, immune antagonism, centrosomal and ciliary disruption, and p53-driven apoptosis collectively explain its central role in ZIKV neurotropism and teratogenicity (Figure 3).
Figure 3
5 Drug research targeting Zika virus-host protein interactions
5.1 Antiviral drugs targeting Zikv-receptor interaction
Currently, there are no licensed antiviral vaccines or medicines for ZIKV infection. R428 (Bemcentinib) is an AXL tyrosine kinase inhibitor, demonstrating dose-dependent reduction of ZIKV infection in human fetal astrocytes and Sertoli cells (). The engineered soluble Axl decoy receptor MYD1 blocks infection by sequestering the bridging ligand Gas6, while polyclonal anti-Axl antibodies significantly reduce infection in endothelial cells and Sertoli cells (). CD300a inhibition targeting Phosphatidylserine receptor has validated as a therapeutic target, with in dendritic cells significantly reducing ZIKV replication (). The lantibiotic peptide duramycin, which binds phosphatidylethanolamine and interferes with PS-mediated entry, inhibited ZIKV replication dose-dependently in astrocytes and Sertoli cells ().
Small-molecule inhibitors targeting the envelope protein have been discovered using high-throughput screening methods—one study screened nearly 27,000 compounds and identified seven that directly block the Zika E protein, thereby preventing viral entry by disrupting its interaction with host cell receptors (; Telehany et al., 2020). Monoclonal antibodies that target unique epitopes on the E protein (including cryptic epitopes on domain III) have also shown potent neutralization, providing an additional strategy to inhibit virus–host attachment and entry (Wu et al., 2017a).
Broad-spectrum entry inhibitors include nanchangmycin, which blocks clathrin-mediated endocytosis and AXL-mediated entry at a pre-fusion stage; suramin, which prevents virus-cell surface binding; and plant-derived compounds such as Aphloia theiformis extracts that impair viral particle attachment. Neutralizing monoclonal antibodies targeting envelope protein epitopes—including C10, 2A10G6 (fusion loop), 5F8 (glycan loop providing full mouse protection), Z23, ZIKV-117 (reducing placental infection and maternal-fetal transmission)—have advanced to Phase 1 clinical trials.
13H10 monoclonal antibodies (anti-ITGB4) not only reduced ZIKV binding and infection in vitro but critically prevented placental infection, reduced embryonic viral titers, and increased embryo survival in pregnant mouse models, demonstrating robust protection against vertical transmission. Heat shock protein inhibitors targeting cell-surface Hsp70 (anti-Hsp70 antibodies and small-molecule inhibitor MKT077) reduced infection and protected mice from lethal ZIKV challenge without inducing resistance. GRP78/BiP antagonism via N-terminus-targeting monoclonal antibodies blocked entry and reduced infection by 50-70%, while siRNA knockdown confirmed its dual role in entry and replication factory formation (). NCAM1 blockade using NCAM1 extracellular domain recombinant protein or anti-NCAM1 antibodies significantly inhibited viral binding and entry, with CRISPR/Cas9 knockout dramatically reducing infection in glioblastoma cells. DC-SIGN antagonism through monoclonal antibodies and the carbohydrate-based blocker mannan prevented viral transmission to target cells. Neuraminidase treatment and genetic deletion of sialic acid biosynthesis genes (GNE, ST3GAL4) greatly reduced infection efficiency across multiple strains, validating the sialic acid pathway as a therapeutic target. Despite extensive preclinical validation, no receptor-targeted antiviral has reached late-stage clinical development, primarily due to receptor redundancy across tissues, cell-type-specific entry mechanisms, and the high safety bar required for pregnancy use.
5.2 Antiviral drugs targeting structural proteins-host proteins interaction
The FDA-approved VCP inhibitor CB-5083 impairs ZIKV infection in both human and mosquito cells and extends survival in flavivirus-infected mice (). Additionally, SERTAD3 promotes capsid degradation, suggesting that enhancing this endogenous antiviral mechanism could suppress viral replication (Sun et al., 2023). Lipid droplet interactions are critical for capsid-mediated nucleocapsid assembly, and lipid metabolism inhibitors such as myriocin (a sphingolipid biosynthesis inhibitor) block ZIKV infection by disrupting lipid droplet-dependent particle assembly ().
Antiviral strategies targeting prM-host protein interactions work by disrupting key proviral pathways through three main mechanisms. First, inhibition of PIM1 kinase reduces phosphorylation of prM at Ser101 and Thr107, which restores AMFR-mediated ubiquitination and degradation of prM. Second, blockade of the cholesterol-binding motifs CARC2 and CARC3, or inhibition of upstream cholesterol biosynthesis, impairs both prM-dependent viral entry and virion assembly. Third, interference with prM folding and maturation through targeting of the ER chaperone GRP78/BiP or the furin protease prevents the cleavage of prM in the trans-Golgi network that normally generates infectious virions ().
5.3 Antiviral drugs targeting non-structural proteins-host proteins interaction
Monoclonal antibodies MAbs 3G2 and 4B8 target ZIKV NS1 and provide protection through Fcγ receptor dependent and independent mechanisms while avoiding antibody dependent enhancement (Yu et al., 2021). Regorafenib, a Raf kinase inhibitor, blocks viral translation, egress, and NS1 secretion to reduce pathogenesis (Wilken et al., 2024). Sofosbuvir, an FDA approved nucleoside analog polymerase inhibitor, suppresses NS5 replication in human cell lines and protects mice from lethal infection (). Galidesivir(BCX4430), a broad spectrum adenosine analog, abrogated viremia in rhesus macaques with favorable CNS penetration ().
Proteasome inhibitors like MG132 block NS2A-mediated degradation of STAT1 and STAT2. This restores interferon signaling and antiviral immunity by directly targeting the NS2A-STAT pathway (). NS3 protease inhibitors prevent proper NS2A maturation by blocking polyprotein processing. Theaflavin-3,3’-digallate is one example that indirectly reduces functional NS2A levels and its pathogenic host protein interactions (). Sofosbuvir reduces overall nonstructural protein expression including NS2A. Raf kinase inhibitors such as Dabrafenib and Regorafenib impair ZIKV replication through post-entry mechanisms. These drugs suppress viral translation and egress and thereby limit NS2A-dependent virion assembly. AMFR E3 ubiquitin ligase inhibitors and FAM134B stabilizers represent emerging therapeutic candidates. These compounds could specifically block the NS2A K56 ubiquitination pathway to restore ER-phagy and prevent congenital pathogenesis.
Doxycycline binds the NS2B-NS3 active site and represents the only protease inhibitor clinically evaluated (). Compound 71 demonstrates superior cellular efficacy as a competitive inhibitor awaiting clinical development. Allosteric inhibitors including SYC-1307, Compounds 1 and 2, temoporfin, and methylene blue target the protease’s “super-open” conformation (; ; ). These drugs show potent activity in enzymatic assays and mouse models by preventing viral protein synthesis and brain damage. Peptide-based inhibitors like C-terminal hexapeptides containing YRRR motifs achieve high catalytic efficiency by occupying substrate-binding sites. Macrocyclic peptides incorporating d-lysine also demonstrate this mechanism ().
Lipophilic statins block cholesterol biosynthesis to impair ZIKV replication that NS4B-DHCR7 interactions facilitate. Atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, and simvastatin reduce viral production by inhibiting HMG-CoA reductase (). Ezetimibe inhibits cholesterol absorption and demonstrates synergistic effects with atorvastatin against ZIKV through complementary mechanisms (). Avasimibe and other SOAT1 inhibitors block cholesterol esterification to impair ZIKV morphogenesis by accumulating free cholesterol and reducing extracellular viral RNA and envelope protein levels (). SBI-0090799 blocks de novo formation of membranous replication compartments by directly targeting the N-terminal region of NS4A (). Dabrafenib and Regorafenib impair ZIKV replication through distinct post-entry mechanisms that affect NS4A and NS4B-dependent viral translation and egress (Wilken et al., 2024). PKI 14–22 suppresses ZIKV infection in endothelial cells and astrocytes by inhibiting protein kinase A without appreciable cytotoxicity (). AGC kinase inhibitors regulate STING signaling through SGK-dependent and independent mechanisms to counteract NS4B-mediated suppression of interferon responses ().
Galidesivir (BCX4430) targets the NS5 RNA-dependent RNA polymerase domain and has completed Phase I clinical trials with demonstrated safety and favorable CNS penetration (). Sofosbuvir inhibits NS5 polymerase activity and protects mice from lethal ZIKV infection while preventing vertical transmission in pregnancy models (). Non-nucleoside RdRp inhibitors bind to the allosteric N pocket and priming loop sites on NS5 polymerase and demonstrate nanomolar potency against ZIKV replication (). AT-9010 binds both the GTP site of NS5 methyltransferase and acts as a chain terminator during RNA synthesis (). SUMOylation inhibitors disrupt NS5 nuclear localization and impair formation of nuclear bodies required for persistent infection (Zhu et al., 2019). CB-5083 inhibits VCP protein function and disrupts NS5-dependent replication organelle formation in both human and mosquito cells (). Suramin blocks NS5-NS3 protein interactions and prevents virus attachment to host cells with pan-flavivirus activity (Yang et al., 2022). The diverse therapeutic strategies targeting ZIKV-host protein interactions span from entry inhibition through replication suppression, with development stages ranging from preclinical evaluation to Phase I clinical trials (Table 2).
Table 2
| Drug | Viral or host target | Development status | Clinical trial stages |
|---|---|---|---|
| Small-molecule E inhibitors | E protein (E domain binding sites) | In vitro screening | None |
| Neutralizing mAbs (C10, 2A10G6, 5F8,Z23, ZIKV-117) | E protein (E protein epitopes) | Phase I clinical trial | Phase I (ongoing) |
| Nanchangmycin;Suramin | E protein entry | Preclinical in vitro/vivo | None |
| R428 (Bemcentinib), MYD1 (AXL decoy receptor); Anti-ITGB4 mAbs (13H10),Recombinant ITGB4; Anti-Hsp70 mAbs,MKT077; Anti-GRP78 mAbs, GRP78 siRNA; NCAM1 extracellular domain; Anti-DC-SIGN mAbs, Mannan carbohydrate; Neuraminidase/ST3GAL4 inhibitors | E protein (AXL pathway; ITGB4 pathway; Hsp70 pathway; GRP78 pathway; NCAM1 pathway; DC-SIGN pathway; sialic acid pathway) | Preclinical in vivo | None |
| CB-5083 | Capsid (VCP ATPase) | Preclinical in vivo | None |
| Statins | prM (Cholesterol biosynthesis) | Preclinical in vitro/vivo | None |
| Proteasome inhibitors; AMFR/FAM134B stabilizers | NS2A (STAT1/STAT2 degradation; FAM134B ubiquitination) | Preclinical in vitro/vivo | None |
| Doxycycline | NS2B-NS3 | Phase I clinical trial | Phase I |
| Compound 71; Allosteric inhibitors; Septin-2 stabilizers | NS2B-NS3 (NS3 protease active site; NS3 protease allosteric site; Septin-2 GTPase) | Preclinical in vitro/vivo | None |
| ANKLE2 stabilizers | NS4A (ANKLE2 ER protein) | Preclinical in vitro | None |
| DHCR7 inhibitors; PKI 14-22 | NS4B (DHCR7 enzyme; TBK1 kinase) | Preclinical in vitro/vivo | None |
| Galidesivir (BCX4430) | NS5 (NS5 RdRp domain) | Phase I clinical trial (completed) | Phase I (completed 2024) |
| Sofosbuvir | NS5 (NS5 RdRp domain) | Preclinical in vitro/vivo | Preclinical |
| Non-nucleoside RdRp inhibitors; AT-9010; SUMOylation inhibitors; p53 activators | NS5(NS5 RdRp allosteric site; NS5 MTase domain; NS5 nuclear impor; p53 tumor suppressort) | Preclinical in vitro | None |
ZIKV therapeutic targets and drug development status.
6 Conclusion and future perspectives
ZIKV enters cells through sequential engagement of multiple receptor classes. ZIKV first binds adhesion receptors including DC-SIGN and Hsp70 for initial attachment, then utilizes high-affinity entry receptors such as ITGB4, GRP78, and NCAM1 that determine cell-type-specific infection. Internalization receptors including integrin αvβ5 and 2,3-linked sialic acid facilitate endocytic uptake, while endosomal receptors such as AXL and TIM-1 mediate low-pH fusion. This multi-receptor strategy enables ZIKV to productively infect neural progenitor cells, placental trophoblasts, and immune cells. Following entry, ZIKV structural proteins including capsid and prM direct virion assembly, while nonstructural proteins NS1-NS5 manipulate immune responses, endoplasmic reticulum architecture, cell-cell junctions, and transcriptional regulation. Although NS5 nuclear localization is shared among flaviviruses, ZIKV NS5 distinctly suppresses neurodevelopmental genes and causes ciliopathy through chromatin binding and PAF1 complex sequestration ().
Identification of these virus-host interactions has revealed multiple therapeutic targets, including receptor antagonists, VCP and protease inhibitors, and NS5 polymerase inhibitors. However, clinical development faces obstacles: functional redundancy allows ZIKV to use alternative entry pathways when single receptors are blocked, multifunctional viral proteins compensate for inhibition of individual functions, and drugs must penetrate the blood-brain and placental barriers while meeting strict pregnancy safety standards. Combination strategies targeting multiple viral proteins simultaneously may overcome these challenges.
ZIKV-host protein interactions are largely conserved across flaviviruses including dengue, West Nile, and Japanese encephalitis viruses. Broad-spectrum antiflaviviral agents would address multiple viral threats, particularly as mosquito vector ranges expand with climate change. Advanced technologies including structural biology, CRISPR screening, and high-throughput screening provide tools for identifying and validating therapeutic targets. Comprehensive knowledge of virus-host interactions will enable rational design of therapeutics capable of preventing ZIKV-induced congenital disease and mitigating future flavivirus epidemics.
Statements
Author contributions
XH: Data curation, Formal analysis, Investigation, Supervision, Validation, Visualization, Writing – original draft, Writing – review & editing. JD: Conceptualization, Data curation, Investigation, Methodology, Validation, Visualization, Writing – original draft, Writing – review & editing. WL: Formal analysis, Methodology, Software, Writing – original draft. SY: Data curation, Formal analysis, Software, Writing – original draft. HS: Conceptualization, Data curation, Software, Writing – original draft. GW: Funding acquisition, Methodology, Project administration, Resources, Supervision, Validation, Visualization, Writing – original draft, Writing – review & editing. HC: Funding acquisition, Project administration, Supervision, Validation, Visualization, Writing – original draft, Writing – review & editing.
Funding
The author(s) declared that financial support was received for this work and/or its publication. This work was supported by the Basic Scientific Research Funds for Inner Mongolia Autonomous Region Government-Directly Affiliated Universities- “Four Prestigious Journals” Landmark Papers “Open Bidding and Leadership Initiative” Project (BR251404), and the General Administration of Customs Project (Grant 2024HK153).
Conflict of interest
The author(s) declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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Summary
Keywords
antiviral drug development, flavivirus pathogenesis, viral entry receptors, virus-host protein interactions, Zika virus
Citation
Han X, Du J, Li W, Yang S, Sun H, Wang G and Cong H (2026) Zika virus and host protein interactions for understanding molecular mechanisms of pathogenesis and therapeutic development. Front. Cell. Infect. Microbiol. 16:1754465. doi: 10.3389/fcimb.2026.1754465
Received
26 November 2025
Revised
05 January 2026
Accepted
08 January 2026
Published
04 February 2026
Volume
16 - 2026
Edited by
Gustavo Ramirez-Martínez, National Institute of Respiratory Diseases-Mexico (INER), Mexico
Reviewed by
Walter Orlando Beys-da-Silva, Federal University of Rio Grande do Sul, Brazil
Jingzhe Shang, Chinese Academy of Medical Sciences, China
Updates
Copyright
© 2026 Han, Du, Li, Yang, Sun, Wang and Cong.
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.
*Correspondence: Haolong Cong, conghaolong@126.com; Guihua Wang, wghua@imau.edu.cn
† These authors have contributed equally to this work
Disclaimer
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