Genetic mapping indicates that VP4 is the rotavirus cell attachment protein in vitro and in vivo

Ludert, Juan E.; Feng, Ningguo; Yu, Jie; Broome, Rosemary L.; Hoshino, Yasutaka; Greenberg, Harry B.

doi:10.1128/jvi.70.1.487-493.1996

Cited by 108 publications

(45 citation statements)

References 40 publications

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“…We provide evidence that the HAstV CP spike is a receptor-binding domain. The CP spike is the outermost domain on the HAstV surface, and the spikes of many other nonenveloped viruses are receptor-binding domains (32)(33)(34). Here, we show that GFP-Spike1, GFP-Spike2, and GFP-Spike8 bind to Caco-2 cells, supporting that all canonical HAstV serotypes may use a common mechanism through the CP spike domain to bind to human cells.…”

Section: Discussionsupporting

confidence: 57%

Structure of a Human Astrovirus Capsid-Antibody Complex and Mechanistic Insights into Virus Neutralization

Bogdanoff

Campos

Perez

et al. 2017

J Virol

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Human astroviruses (HAstVs) are a leading cause of viral diarrhea in young children, the immunocompromised, and the elderly. There are no vaccines or antiviral therapies against HAstV disease. Several lines of evidence point to the presence of protective antibodies in healthy adults as a mechanism governing protection against reinfection by HAstV. However, development of anti-HAstV therapies is hampered by the gap in knowledge of protective antibody epitopes on the HAstV capsid surface. Here, we report the structure of the HAstV capsid spike domain bound to the neutralizing monoclonal antibody PL-2. The antibody uses all six complementarity-determining regions to bind to a quaternary epitope on each side of the dimeric capsid spike. We provide evidence that the HAstV capsid spike is a receptor-binding domain and that the antibody neutralizes HAstV by blocking virus attachment to cells. We identify patches of conserved amino acids that overlap the antibody epitope and may comprise a receptor-binding site. Our studies provide a foundation for the development of therapies to prevent and treat HAstV diarrheal disease.IMPORTANCE Human astroviruses (HAstVs) infect nearly every person in the world during childhood and cause diarrhea, vomiting, and fever. Despite the prevalence of this virus, little is known about how antibodies in healthy adults protect them against reinfection. Here, we determined the crystal structure of a complex of the HAstV capsid protein and a virus-neutralizing antibody. We show that the antibody binds to the outermost spike domain of the capsid, and we provide evidence that the antibody blocks virus attachment to human cells. Importantly, our findings suggest that a subunit-based vaccine focusing the immune system on the HAstV capsid spike domain could be effective in protecting children against HAstV disease.KEYWORDS antibody, astrovirus, capsid, receptor binding, structure, vaccines, virus neutralization T he Astroviridae family is comprised of two genera, Mamastrovirus and Avastrovirus, which infect mammalian and avian species, respectively (1). Members of the Avastrovirus genus cause a variety of disease manifestations, growth defects, and mortality in poultry (2). Members of the Mamastrovirus genus cause infections in humans and a wide range of mammals (3). Human astroviruses (HAstVs) are classified into eight canonical serotypes (HAstV-1 to ) within the Mamastrovirus genogroup 1 (4), where HAstV-1 is the predominant serotype worldwide (5, 6). HAstV is a leading cause of viral diarrhea in children, immunocompromised individuals, and the elderly (7). There are approximately 3.9 million cases of viral diarrhea due to HAstV in the United States every year (8). In addition, highly divergent strains of HAstV have recently been attributed to encephalitis in immunocompromised individuals (9-11).

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Section: Discussionsupporting

confidence: 57%

Structure of a Human Astrovirus Capsid-Antibody Complex and Mechanistic Insights into Virus Neutralization

Bogdanoff

Campos

Perez

et al. 2017

J Virol

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“…3B). The binding of RRV to cells treated with an siRNA against CD-M6PR (siCD-M6PR) was not significantly affected compared to that in control cells transfected with an irrelevant siRNA, while it was severely decreased when the CD-M6PR-knockdown cells were treated with neuraminidase (NA), which removes the sialic acid required for cell attachment of RRV (54). In contrast, the binding of the UK virus was not affected in CD-M6PR-knockdown cells treated or not with NA ( Fig.…”

Section: Rotavirus Strains Uk and Rrv Reach Different Endocytic Compamentioning

confidence: 99%

Rotaviruses Reach Late Endosomes and Require the Cation-Dependent Mannose-6-Phosphate Receptor and the Activity of Cathepsin Proteases To Enter the Cell

Díaz-Salinas

Silva‐Ayala

López

2014

J Virol

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Rotaviruses (RVs) enter cells through different endocytic pathways. Bovine rotavirus (BRV) UK uses clathrin-mediated endocytosis, while rhesus rotavirus (RRV) employs an endocytic process independent of clathrin and caveolin. Given the differences in the cell internalization pathway used by these viruses, we tested if the intracellular trafficking of BRV UK was the same as that of RRV, which is known to reach maturing endosomes (MEs) to infect the cell. We found that BRV UK also reaches MEs, since its infectivity depends on the function of Rab5, the endosomal sorting complex required for transport (ESCRT), and the formation of endosomal intraluminal vesicles (ILVs). However, unlike RRV, the infectivity of BRV UK was inhibited by knocking down the expression of Rab7, indicating that it has to traffic to late endosomes (LEs) to infect the cell. The requirement for Rab7 was also shared by other RV strains of human and porcine origin. Of interest, most RV strains that reach LEs were also found to depend on the activities of Rab9, the cation-dependent mannose-6-phosphate receptor (CD-M6PR), and cathepsins B, L, and S, suggesting that cellular factors from the trans-Golgi network (TGN) need to be transported by the CD-M6PR to LEs to facilitate RV cell infection. Furthermore, using a collection of UK ؋ RRV reassortant viruses, we found that the dependence of BRV UK on Rab7, Rab9, and CD-M6PR is associated with the spike protein VP4. These findings illustrate the elaborate pathway of RV entry and reveal a new process (Rab9/CD-M6PR/cathepsins) that could be targeted for drug intervention. IMPORTANCERotavirus is an important etiological agent of severe gastroenteritis in children. In most instances, viruses enter cells through an endocytic pathway that delivers the viral particle to vesicular organelles known as early endosomes (EEs). Some viruses reach the cytoplasm from EEs, where they start to replicate their genome. However, other viruses go deeper into the cell, trafficking from EEs to late endosomes (LEs) to disassemble and reach the cytoplasm. In this work, we show that most RV strains have to traffic to LEs, and the transport of endolysosomal proteases from the Golgi complex to LEs, mediated by the mannose-6-phosphate receptor, is necessary for the virus to exit the vesicular compartment and efficiently start viral replication. We also show that this deep journey into the cell is associated with the virus spike protein VP4. These findings illustrate the elaborate pathway of RV entry that could be used for drug intervention.

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“…11 The attachment of the virus to cell surface receptors is mediated by VP4 (88 kDa). 12 Trypsin cleavage of VP4 generates two virion-associated polypeptides: VP5* (60 kDa) and VP8* (28 kDa). This event greatly enhances viral infectivity, induces membrane permeability, and is necessary for maximal viral growth.…”

Section: Introductionmentioning

confidence: 99%

Localization of membrane permeabilization and receptor binding sites on the VP4 hemagglutinin of rotavirus: implications for cell entry

Tihova

Dryden

Bellamy

et al. 2001

Journal of Molecular Biology

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Genetic mapping indicates that VP4 is the rotavirus cell attachment protein in vitro and in vivo

Cited by 108 publications

References 40 publications

Structure of a Human Astrovirus Capsid-Antibody Complex and Mechanistic Insights into Virus Neutralization

Structure of a Human Astrovirus Capsid-Antibody Complex and Mechanistic Insights into Virus Neutralization

Rotaviruses Reach Late Endosomes and Require the Cation-Dependent Mannose-6-Phosphate Receptor and the Activity of Cathepsin Proteases To Enter the Cell

Localization of membrane permeabilization and receptor binding sites on the VP4 hemagglutinin of rotavirus: implications for cell entry

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