Amino acid domains 280–297 of VP6 and 531–554 of VP4 are implicated in heat shock cognate protein hsc70-mediated rotavirus infection

Gualtero, Diego; Guzmán, Fanny; Acosta, Orlando; Guerrero, C.

doi:10.1007/s00705-007-1055-5

Cited by 36 publications

(44 citation statements)

References 41 publications

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“…Their binding could block VP6 interaction with a cellular receptor or induce a conformational change in the viral particle, interfering with virus attachment to the cell. This hypothesis is supported by previous data demonstrating that VP4 and VP6 bind the hsp70 cellular protein (22). On the other hand, if the VHH-virus complex can attach and enter the cell, virus decapsidation or transcriptional activity and mRNA exit might be blocked.…”

Section: Discussionsupporting

confidence: 77%

Llama-Derived Single-Chain Antibody Fragments Directed to Rotavirus VP6 Protein Possess Broad Neutralizing Activity In Vitro and Confer Protection against Diarrhea in Mice

Garaicoechea¹,

Olichon

Marcoppido³

et al. 2008

J Virol

100

105

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Group A rotavirus is one of the most common causes of severe diarrhea in human infants and newborn animals. Rotavirus virions are triple-layered particles. The outer capsid proteins VP4 and VP7 are highly variable and represent the major neutralizing antigens. The inner capsid protein VP6 is conserved among group A rotaviruses, is highly immunogenic, and is the target antigen of most immunodiagnosis tests. Llama-derived single-chain antibody fragments (VHH) are the smallest molecules with antigen-binding capacity and can therefore be expected to have properties different from conventional antibodies. In this study a library containing the VHH genes of a llama immunized with recombinant inner capsid protein VP6 was generated. Binders directed to VP6, in its native conformation within the viral particle, were selected and characterized. Four selected VHH directed to conformational epitopes of VP6 recognized all human and animal rotavirus strains tested and could be engineered for their use in immunodiagnostic tests for group A rotavirus detection. Three of the four VHH neutralized rotavirus in vivo independently of the strain serotype. Furthermore, this result was confirmed by in vivo partial protection against rotavirus challenge in a neonatal mouse model. The present study demonstrates for the first time a broad neutralization activity of VP6 specific VHH in vitro and in vivo. Neutralizing VHH directed to VP6 promise to become an essential tool for the prevention and treatment of rotavirus diarrhea.Group A rotavirus (RV) is the leading cause of acute gastroenteritis in human infants less than 5 years old, causing 611,000 deaths per year (41). It is also the main cause of severe diarrhea in the neonates of many animal species of economic interest (43,47). RV virions are triple-layered particles composed by a core (protein VP2), an inner capsid (protein VP6), and an outer capsid (proteins VP7 and VP4) (16,29). The inner capsid protein, VP6, is a trimer representing 51% of the virion mass. According to the antigenic variation of VP6, RVs are classified into seven groups (A to G) (16). Depending on the presence or absence of two different epitopes in the VP6 protein, group A RV strains are further divided into subgroups (Sb) I, II, IϩII, and no I no II. Despite the different subgroups mentioned, VP6 is a strongly conserved protein among all group A RVs (Ͼ90% amino acid homology). It is highly immunogenic and constitutes the target antigen of most immunodiagnosis tests for group A RV detection. In contrast, the outer capsid proteins VP7 (glycoprotein) and VP4 (protease sensitive) are highly variable and constitute the major neutralizing antigens. Based on the variation of VP7 and VP4, group A RVs are further classified into G and P types, respectively.

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Section: Discussionsupporting

confidence: 77%

Llama-Derived Single-Chain Antibody Fragments Directed to Rotavirus VP6 Protein Possess Broad Neutralizing Activity In Vitro and Confer Protection against Diarrhea in Mice

Garaicoechea¹,

Olichon

Marcoppido³

et al. 2008

J Virol

100

105

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“…Las células se incubaron a 37 °C hasta la lisis celular; se recuperó el lisado y se almacenó a -20 °C hasta su uso. El lisado celular fue congelado y descongelado dos veces y titulado por su actividad infecciosa en términos de unidades formadoras de foco (21).…”

Section: Propagación De Las Cepas De Rotavirusunclassified

“…Se considera que los rotavirus utilizan como receptor funcional un complejo de varias macromoléculas que incluyen, entre otras moléculas, las integrinas αvβ3, α2β1, αxβ2 y la proteína de choque térmico Hsc70, cuya presencia se ha evidenciado especialmente en la superficie de células MA104 (8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23). Recientemente, en un estudio realizado en nuestro laboratorio (Calderón MN, Acosta O, Guerrero CA, Guzmán F. Protein disulfide isomerase activity is involved in rotavirus entry to MA104 cells.…”

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“…2008. p. 163-4), se encontró que bloqueadores de la actividad de la proteína disulfuro-isomerasa de la superficie celular y anticuerpos anti-proteína disulfuro-isomerasa inhiben la infección por rotavirus en células MA104. Estos hallazgos sugieren una mayor complejidad en el mecanismo de entrada del rotavirus a la célula y la existencia de posibles vías alternativas para la entrada de las diferentes cepas de rotavirus a las células huésped (14,15,21,24).…”

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Interacción de rotavirus con la proteína disulfuro-isomerasa in vitro y en sistemas celulares

et al. 2011

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“…Histoblood group antigens bound by VP8* also are implicated as cellular receptors for certain rotaviruses (21,22). Additional proposed entry factors include ␣x␤2 and ␣v␤3 integrins, heat shock cognate protein 70, and lipid membrane microdomains sensitive to cholesterol depletion (8,15,(23)(24)(25)(26). Naturally occurring mammalian rotaviruses exhibit various abilities to utilize Sia, gangliosides, and integrins (7,14,23,(27)(28)(29)(30)(31).…”

mentioning

confidence: 99%

Relative Roles of GM1 Ganglioside, N -Acylneuraminic Acids, and α2β1 Integrin in Mediating Rotavirus Infection

Fleming

Böhm

Dang

et al. 2014

J Virol

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N-acetyl-and N-glycolylneuraminic acids (Sia) and ␣2␤1 integrin are frequently used by rotaviruses as cellular receptors through recognition by virion spike protein VP4. The VP4 subunit VP8*, derived from Wa rotavirus, binds the internal N-acetylneuraminic acid on ganglioside GM1. Wa infection is increased by enhanced internal Sia access following terminal Sia removal from main glycan chains with sialidase. The GM1 ligand cholera toxin B (CTB) reduces Wa infectivity. Here, we found sialidase treatment increased cellular GM1 availability and the infectivity of several other human (including RV-3) and animal rotaviruses, typically rendering them susceptible to methyl ␣-D-N-acetylneuraminide treatment, but did not alter ␣2␤1 usage. CTB reduced the infectivity of these viruses. Aceramido-GM1 inhibited Wa and RV-3 infectivity in untreated and sialidasetreated cells, and GM1 supplementation increased their infectivity, demonstrating the importance of GM1 for infection. Wa recognition of ␣2␤1 and internal Sia were at least partially independent. Rotavirus usage of GM1 was mapped to VP4 using virus reassortants, and RV-3 VP8* bound aceramido-GM1 by saturation transfer difference nuclear magnetic resonance (STD NMR). Most rotaviruses recognizing terminal Sia did not use GM1, including RRV. RRV VP8* interacted minimally with aceramido-GM1 by STD NMR. Unusually, TFR-41 rotavirus infectivity depended upon terminal Sia and GM1. Competition of CTB, Sia, and/or aceramido-GM1 with cell binding by VP8* from representative rotaviruses showed that rotavirus Sia and GM1 preferences resulted from VP8*-cell binding. Our major finding is that infection by human rotaviruses of commonly occurring VP4 serotypes involves VP8* binding to cell surface GM1 glycan, typically including the internal N-acetylneuraminic acid. IMPORTANCERotaviruses, the major cause of severe infantile gastroenteritis, recognize cell surface receptors through virus spike protein VP4. Several animal rotaviruses are known to bind sialic acids at the termini of main carbohydrate chains. Conversely, only a single human rotavirus is known to bind sialic acid. Interestingly, VP4 of this rotavirus bound to sialic acid that forms a branch on the main carbohydrate chain of the GM1 ganglioside. Here, we use several techniques to demonstrate that other human rotaviruses exhibit similar GM1 usage properties. Furthermore, binding by VP4 to cell surface GM1, involving branched sialic acid recognition, is shown to facilitate infection. In contrast, most animal rotaviruses that bind terminal sialic acids did not utilize GM1 for VP4 cell binding or infection. These studies support a significant role for GM1 in mediating host cell invasion by human rotaviruses.

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Amino acid domains 280–297 of VP6 and 531–554 of VP4 are implicated in heat shock cognate protein hsc70-mediated rotavirus infection

Cited by 36 publications

References 41 publications

Llama-Derived Single-Chain Antibody Fragments Directed to Rotavirus VP6 Protein Possess Broad Neutralizing Activity In Vitro and Confer Protection against Diarrhea in Mice

Llama-Derived Single-Chain Antibody Fragments Directed to Rotavirus VP6 Protein Possess Broad Neutralizing Activity In Vitro and Confer Protection against Diarrhea in Mice

Interacción de rotavirus con la proteína disulfuro-isomerasa in vitro y en sistemas celulares

Relative Roles of GM1 Ganglioside, N -Acylneuraminic Acids, and α2β1 Integrin in Mediating Rotavirus Infection

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