The Envelope Residues E152/156/158 of Zika Virus Influence the Early Stages of Virus Infection in Human Cells

Emerging infections of mosquito-borne Zika virus (ZIKV) pose an increasing threat to human health, as documented over the recent years in South Pacific islands and the Americas in recent years. To better understand molecular mechanisms underlying the increase in human cases with severe pathologies, we recently demonstrated the functional roles of structural proteins capsid (C), pre-membrane (prM), and envelop (E) of ZIKV epidemic strains with the initiation of viral infection in human cells. Specifically, we found that the C-prM region contributes to permissiveness of human host cells to ZIKV infection and ZIKV-induced cytopathic effects, whereas the E protein is associated with viral attachment and early infection. In the present study, we further characterize ZIKV E proteins by investigating the roles of residues isoleucine 152 (Ile152), threonine 156 (Thr156), and histidine 158 (His158) (i.e., the E-152/156/158 residues), which surround a unique N-glycosylation site (E-154), in permissiveness of human host cells to epidemic ZIKV infection. For comparison purpose, we generated mutant molecular clones of epidemic BeH819015 (BR15) and historical MR766-NIID (MR766) strains that carry each other’s E-152/156/158 residues, respectively. We observed that the BR15 mutant containing the E-152/156/158 residues from MR766 was less infectious in A549-Dual™ cells than parental virus. In contrast, the MR766 mutant containing E-152/156/158 residues from BR15 displayed increased infectivity. The observed differences in infectivity were, however, not correlated with changes in viral binding onto host-cells or cellular responses to viral infection. Instead, the E-152/156/158 residues from BR15 were associated with an increased efficiency of viral membrane fusion inside infected cells due to conformational changes of E protein that enhance exposure of the fusion loop. Our data highlight an important contribution of E-152/156/158 residues to the early steps of ZIKV infection in human cells.

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High-throughput fitness profiling of Zika virus E protein reveals different roles for N-linked glycosylation during infection of mammalian and mosquito cells

10.1101/199935 ◽

2017 ◽

Author(s):

Danyang Gong ◽

Tian-hao Zhang ◽

Dawei Zhao ◽

Yushen Du ◽

Travis J. Chapa ◽

...

Keyword(s):

Zika Virus ◽

Cell Receptor ◽

Human Cells ◽

Nucleotide Position ◽

List Type ◽

Zikv Infection ◽

E Protein ◽

Mosquito Cells ◽

Viral Mutant ◽

Replication Fitness

AbstractZika virus (ZIKV) infection causes Guillain-Barré syndrome and severe birth defects. ZIKV envelope (E) protein is the major viral protein involved in cell receptor binding and entry and therefore considered one of the major determinants in ZIKV pathogenesis. Here, we report a gene-wide mapping of functional residues of ZIKV E protein using a mutant library with changes covering every nucleotide position. By comparing the replication fitness of every viral mutant between mosquito and human cells, we identified that mutations affecting N-linked glycosylation at N154 position display the most divergence. Through characterizing individual mutants, we show that, while ablation of N-linked glycosylation selectively benefits ZIKV infection of mosquito cells by enhancing cell entry, it either had little impact on ZIKV infection on certain human cells or decreased infection through entry factor DC-SIGN. In conclusion, we define the roles of individual residues of ZIKV envelope protein, which contribute to ZIKV replication fitness in human and mosquito cells.HighlightsGene-wide mapping of functional residues of E protein in human and mosquito cells.Mutations affecting N-linked glycosylation display the most dramatic difference.N-linked glycosylation decreases ZIKV entry into mosquito cells.N-linked glycosylation is important for DC-SIGN mediated infection of human cells.

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Corrigendum to “The structural proteins of epidemic and historical strains of Zika virus differ in their ability to initiate viral infection in human host cells” [Virology 516 (2018) 265–273]

Virology ◽

10.1016/j.virol.2018.11.006 ◽

2019 ◽

Vol 526 ◽

pp. 233

Author(s):

Sandra Bos ◽

Wildriss Viranaicken ◽

Jonathan Turpin ◽

Chaker El-Kalamouni ◽

Marjolaine Roche ◽

...

Keyword(s):

Viral Infection ◽

Zika Virus ◽

Host Cells ◽

Structural Proteins ◽

Human Host

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The structural proteins of epidemic and historical strains of Zika virus differ in their ability to initiate viral infection in human host cells

Virology ◽

10.1016/j.virol.2017.12.003 ◽

2018 ◽

Vol 516 ◽

pp. 265-273 ◽

Cited By ~ 25

Author(s):

Sandra Bos ◽

Wildriss Viranaicken ◽

Jonathan Turpin ◽

Chaker El-Kalamouni ◽

Marjolaine Roche ◽

...

Keyword(s):

Viral Infection ◽

Zika Virus ◽

Host Cells ◽

Structural Proteins ◽

Human Host

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Ayapana triplinervis Essential Oil and Its Main Component Thymohydroquinone Dimethyl Ether Inhibit Zika Virus at Doses Devoid of Toxicity in Zebrafish

Molecules ◽

10.3390/molecules24193447 ◽

2019 ◽

Vol 24 (19) ◽

pp. 3447 ◽

Cited By ~ 5

Author(s):

Juliano G. Haddad ◽

Morgane Picard ◽

Sebastien Bénard ◽

Claire Desvignes ◽

Philippe Desprès ◽

...

Keyword(s):

Medicinal Plants ◽

Acute Toxicity ◽

Essential Oil ◽

Dimethyl Ether ◽

Zika Virus ◽

Human Cells ◽

Reunion Island ◽

Zikv Infection ◽

Réunion Island ◽

Main Component

Zika virus (ZIKV) is an emerging mosquito-borne virus of medical concern. ZIKV infection may represent a serious disease, causing neonatal microcephaly and neurological disorders. Nowadays, there is no approved antiviral against ZIKV. Several indigenous or endemic medicinal plants from Mascarene archipelago in Indian Ocean have been found able to inhibit ZIKV infection. The purpose of our study was to determine whether essential oil (EO) from Reunion Island medicinal plant Ayapana triplinervis, whose thymohydroquinone dimethyl ether (THQ) is the main component has the potential to prevent ZIKV infection in human cells. Virological assays were performed on human epithelial A549 cells infected with either GFP reporter ZIKV or epidemic viral strain. Zebrafish assay was employed to evaluate the acute toxicity of THQ in vivo. We showed that both EO and THQ inhibit ZIKV infection in human cells with IC50 values of 38 and 45 µg/mL, respectively. At the noncytotoxic concentrations, EO and THQ reduced virus progeny production by 3-log. Time-of-drug-addition assays revealed that THQ could act as viral entry inhibitor. At the antiviral effective concentration, THQ injection in zebrafish does not lead to any signs of stress and does not impact fish survival, demonstrating the absence of acute toxicity for THQ. From our data, we propose that THQ is a new potent antiviral phytocompound against ZIKV, supporting the potential use of medicinal plants from Reunion Island as a source of natural and safe antiviral substances against medically important mosquito-borne viruses.

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Sulfated Glycans and Related Digestive Enzymes in the Zika Virus Infectivity: Potential Mechanisms of Virus-Host Interaction and Perspectives in Drug Discovery

Interdisciplinary Perspectives on Infectious Diseases ◽

10.1155/2017/4894598 ◽

2017 ◽

Vol 2017 ◽

pp. 1-8 ◽

Cited By ~ 2

Author(s):

Vitor H. Pomin

Keyword(s):

Neural Development ◽

Zika Virus ◽

Molecular Mechanisms ◽

Surface Proteins ◽

Host Cells ◽

Future Research ◽

Virus Infectivity ◽

Virus Attachment ◽

Chemical Structures ◽

Host Interaction

As broadly reported, there is an ongoing Zika virus (ZIKV) outbreak in countries of Latin America. Recent findings have demonstrated that ZIKV causes severe defects on the neural development in fetuses in utero and newborns. Very little is known about the molecular mechanisms involved in the ZIKV infectivity. Potential therapeutic agents are also under investigation. In this report, the possible mechanisms of action played by glycosaminoglycans (GAGs) displayed at the surface proteoglycans of host cells, and likely in charge of interactions with surface proteins of the ZIKV, are highlighted. As is common for the most viruses, these sulfated glycans serve as receptors for virus attachment onto the host cells and consequential entry during infection. The applications of (1) exogenous sulfated glycans of different origins and chemical structures capable of competing with the virus attachment receptors (supposedly GAGs) and (2) GAG-degrading enzymes able to digest the virus attachment receptors on the cells may be therapeutically beneficial as anti-ZIKV. This communication attempts, therefore, to offer some guidance for the future research programs aimed to unveil the molecular mechanisms underlying the ZIKV infectivity and to develop therapeutics capable of decreasing the devastating consequences caused by ZIKV outbreak in the Americas.

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CImP: Cellular Imprinting Proteomics applied to ocular disorders elicited by Congenital Zika virus Syndrome

10.1101/648600 ◽

2019 ◽

Cited By ~ 1

Author(s):

Livia Rosa-Fernandes ◽

Raquel Hora Barbosa ◽

Maria Luiza B. dos Santos ◽

Claudia B. Angeli ◽

Thiago P. Silva ◽

...

Keyword(s):

Zika Virus ◽

Ocular Surface ◽

Molecular Mechanisms ◽

Membrane Filter ◽

Neurological Diseases ◽

Specific Protein ◽

Protein Markers ◽

Impression Cytology ◽

Zikv Infection ◽

Molecular Alterations

AbstractIMPORTANCEOcular complications in infants with Congenital Zika Syndrome (CZS) have been reported. However, the molecular mechanisms underlying of eye dysfunctions are presently unknown.OBJECTIVEA method (termed Cellular Imprinting Proteomics, CImP) for the identification and quantification of the ocular surface proteome using a minimally invasive membrane filter device is described. Moreover, The CImP method was applied to profile the molecular alterations in the eyes of infants exposed to Zika virus (ZIKV) infection during gestation.DESIGN, SETTINGS AND PARTICIMPANTSThe CImP method was applied to a cohort divided into three conditions: 1) Ctrl (infants with no infectious diseases, n=5). 2) Zikv (infants exposed to ZIKV gestation, with no microcephaly, n=5). 3) ZikvCZS(infants exposed to ZIKV, with microcephaly, n=3). All conditions were age and sex-matched. An improved impression cytology method was used to capture the outermost ocular surface cells. The number of impression cytology membrane collected was: Ctrl (12), Zikv (14) and ZikvCZS(8). Proteins were extracted and analysed using mass spectrometry-based proteomics technology followed by statistical analysis. Parallel reaction monitoring was performed to validate the expression of specific protein markers.RESULTSUsing the CImP method, 2209 proteins were identified on the membrane-captured conjunctiva epithelial cells. Modulation of neutrophil degranulation, cell death, ocular and neurodevelopment pathways are reported in infants with CZS compared to matched controls. Moreover, the molecular pattern of ocular surface cells retrieved from infants infected during the gestation but with no CZS was different from matched controls.CONCLUSIONS AND PERSPECTIVESMolecular alterations in the ocular cell surface associated to ZIKV infection with and without CZS complications are reported for the first time. We predict that this method will be introduced successfully in the study of several neurological diseases with the aim to identify novel diagnostic and therapeutic biomarkers.

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A BRIEF REVIEW ON ZIKA VIRUS INFECTION

Brazilian Journal of Medicine and Human Health ◽

10.17267/2317-3386bjmhh.v4i2.961 ◽

2016 ◽

Vol 4 (2) ◽

Author(s):

Maria Fernanda Rios Grassi ◽

Antônio Carlos Albuquerque Bandeira ◽

Luana Leandro Gois ◽

Geraldo Gileno de Sá Oliveira

Keyword(s):

Immune Response ◽

Natural History ◽

Virus Infection ◽

Southeast Asia ◽

Zika Virus ◽

Pacific Islands ◽

Neurological Manifestations ◽

Zikv Infection ◽

The World

Since isolation of Zika virus (ZIKV) in Uganda from Zika forest in the 1947, for sixty years the virus has caused only scattered human cases in Africa and Southeast Asia. From 2007, outbreaks with an increasing number of cases, including cases with neurological manifestations, have been occurring in Pacific islands. In 2015, ZIKV reached Brazil with an explosive number of cases and a severe neurological impact on fetuses and newborns. The natural history and several immunological aspects of ZIKV infection need to be characterized. In this review it is summarized the spread of ZIKV around the world and pointed out some gaps on the immunological knowledge related to the infection. The characterization of the immunodominant/protective immune response would contribute to vaccine and diagnosis tests development.

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Adaptation to host cell environment during experimental evolution of Zika virus

10.21203/rs.3.rs-492894/v1 ◽

2021 ◽

Author(s):

Vincent Grass ◽

Emilie Hardy ◽

Kassian Kobert ◽

Soheil Rastgou Talemi ◽

Elodie Décembre ◽

...

Keyword(s):

Host Cell ◽

Zika Virus ◽

Immune Escape ◽

Viral Evolution ◽

Time Lag ◽

Host Cells ◽

Host Responses ◽

Type I ◽

Zikv Infection ◽

Cellular Environment

Abstract Zika virus (ZIKV) infection can cause important developmental and neurological defects in Humans. Type I/III interferon responses control ZIKV infection and pathological processes, yet the virus has evolved various mechanisms to defeat these host responses. Here, we established a pipeline to delineate at high-resolution the genetic evolution of ZIKV in a controlled host cell environment. We uncovered that serially passaged ZIKV acquired increased infectivity and simultaneously developed a resistance to TLR3-induced restriction. We built a mathematical model that suggests that the increased infectivity is due to a reduced time-lag between infection and viral replication. We found that this adaptation is cell-type specific, suggesting that different cell environments may drive viral evolution along different routes. Deep-sequencing of ZIKV populations pinpointed mutations whose increased frequencies temporally coincide with the acquisition of the adapted phenotype. We functionally validated S455L, a substitution in ZIKV envelope (E) protein, recapitulating the adapted phenotype. Its positioning on the E structure suggests a putative function in protein refolding/stability. Taken together, our results uncovered ZIKV adaptations to the cellular environment leading to accelerated replication onset coupled with resistance to TLR3-induced antiviral response. Our work provides insights into Zika virus adaptation to host cells and immune escape mechanisms.

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ADP-Ribosyltransferase PARP11 Suppresses Zika Virus in Synergy with PARP12

10.21203/rs.3.rs-385154/v1 ◽

2021 ◽

Author(s):

Lili Li ◽

Yueyue Shi ◽

Sirui Li ◽

Junxiao Liu ◽

Shulong Zu ◽

...

Keyword(s):

Zika Virus ◽

Neurological Complications ◽

Host Cells ◽

Interferon Response ◽

Type I ◽

Zikv Infection ◽

Interferon Stimulated Genes ◽

Ns3 Protein ◽

Global Threat ◽

Adp Ribosyltransferase

Abstract Zika virus (ZIKV) infection and ZIKV epidemic have been continuously spreading silently throughout the world and its associated microcephaly and other serious congenital neurological complications poses a significant global threat to public health. ZIKV infection stimulates type I interferon response in host cells which suppresses viral replication by inducing the expression of interferon-stimulated genes (ISGs). Here, we identified ADP-ribosyltransferase PARP11 as an anti-ZIKV ISG and found that PARP11 suppressed ZIKV independently on itself PARP enzyme activity. Furthermore, PARP11 interacted with PARP12 and promoted PARP12-mediating ZIKV NS1 and NS3 protein degradation. Homo family PARP11 and PARP12 cooperated with each other on ZIKV suppression and the anti-ZIKV function of PARP11 mostly dependent on the existence of PARP12. Our findings have broadened the understanding of the anti-viral function of PARP11, and more importantly suggest a potential therapeutics target against ZIKV infection.

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A Novel Mechanism for Zika Virus Host-Cell Binding

Viruses ◽

10.3390/v11121101 ◽

2019 ◽

Vol 11 (12) ◽

pp. 1101

Author(s):

Courtney A. Rieder ◽

Jonathan Rieder ◽

Sebastién Sannajust ◽

Diana Goode ◽

Ramaz Geguchadze ◽

...

Keyword(s):

Zika Virus ◽

Neuronal Cells ◽

Annexin V ◽

Vero Cells ◽

Western Hemisphere ◽

Host Cells ◽

E Protein ◽

Clinical Presentations ◽

Secondary Mechanism ◽

Protein Cell

Zika virus (ZIKV) recently emerged in the Western Hemisphere with previously unrecognized or unreported clinical presentations. Here, we identify two putative binding mechanisms of ancestral and emergent ZIKV strains featuring the envelope (E) protein residue asparagine 154 (ASN154) and viral phosphatidylserine (PS). Synthetic peptides representing the region containing ASN154 from strains PRVABC59 (Puerto Rico 2015) and MR_766 (Uganda 1947) were exposed to neuronal cells and fibroblasts to model ZIKV E protein/cell interactions and bound MDCK or Vero cells and primary neurons significantly. Peptides significantly inhibited Vero cell infectivity by ZIKV strains MR_766 and PRVABC59, indicating that this region represents a putative binding mechanism of ancestral African ZIKV strains and emergent Western Hemisphere strains. Pretreatment of ZIKV strains MR_766 and PRVABC59 with the PS-binding protein annexin V significantly inhibited replication of PRVABC59 but not MR_766, suggesting that Western hemisphere strains may additionally be capable of utilizing PS-mediated entry to infect host cells. These data indicate that the region surrounding E protein ASN154 is capable of binding fibroblasts and primary neuronal cells and that PS-mediated entry may be a secondary mechanism for infectivity utilized by Western Hemisphere strains.

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