Gangliosides Have a Functional Role during Rotavirus Cell Entry

Martı́nez, Miguel Ángel; López, Susana; Arias, Carlos F.; Iša, Pavel

doi:10.1128/jvi.01964-12

Cited by 63 publications

(48 citation statements)

References 45 publications

(52 reference statements)

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“…This is consistent with TFR-41 usage of GM1, which lacks terminal Sia. Our findings that GM1 is used by UK and TFR-41 extend the recent report that suppression of ganglioside synthesis inhibits the infectivity of these rotaviruses (72).…”

Section: Discussionsupporting

confidence: 77%

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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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Section: Discussionsupporting

confidence: 77%

“…Another study has detected GM1 inhibition of RRV infection (72). However, this may relate to the presence of the ceramide moiety, which was absent from our a-GM1, rather than the GM1 glycan itself.…”

Section: Discussionmentioning

confidence: 55%

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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“…Though the binding assay performed in this study may not have discriminated against nonspecific binding of treated or untreated viruses, k= binding rate constants served to indicate the occurrence of binding inhibition in three of the five treatments tested: heat treatment at 57°C and the two solar treatments in which the organic sensitizer was present. Because proteins are the only rotavirus components able to attach to the host receptors, binding inhibition may also indicate inhibition of rotavirus entry, as structural proteins VP4 and VP7 are responsible for both binding and entry (13,37,38). It should be noted that protein conformations have been associated with heat treatment of viruses (21,24).…”

Section: Discussionmentioning

confidence: 99%

Solar and Temperature Treatments Affect the Ability of Human Rotavirus Wa To Bind to Host Cells and Synthesize Viral RNA

Romero‐Maraccini

Shisler

Nguyen

2015

Appl Environ Microbiol

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Rotavirus, the leading cause of diarrheal diseases in children under the age of five, is often resistant to conventional wastewater treatment and thus can remain infectious once released into the aquatic environment. Solar and heat treatments can inactivate rotavirus, but it is unknown how these treatments inactivate the virus on a molecular level. To answer this question, our approach was to correlate rotavirus inactivation with the inhibition of portions of the virus life cycle as a means to identify the mechanisms of solar or heat inactivation. Specifically, the integrity of the rotavirus NSP3 gene, virus-host cell interaction, and viral RNA synthesis were examined after heat (57°C) or solar treatment of rotavirus. Only the inhibition of viral RNA synthesis positively correlated with a loss of rotavirus infectivity; 57°C treatment of rotavirus resulted in a decrease of rotavirus RNA synthesis at the same rate as rotavirus infectivity. These data suggest that heat treatment neutralized rotaviruses primarily by targeting viral transcription functions. In contrast, when using solar disinfection, the decrease in RNA synthesis was responsible for approximately one-half of the decrease in infectivity, suggesting that other mechanisms, including posttranslational, contribute to inactivation. Nevertheless, both solar and heat inactivation of rotaviruses disrupted viral RNA synthesis as a mechanism for inactivation.O f all waterborne microbes that can cause pathogenic diseases, viruses remain the most challenging for study in aquatic environments (1-4). This is for several reasons. First, compared to bacterial pathogens, viruses are more resistant to disinfection by standard water disinfection treatments (2, 5, 6). Second, for those conditions that do indeed neutralize viruses, the molecular mechanism(s) responsible for inactivation is unclear (7,8). Third, it is only recently that technologies such as quantitative PCR (qPCR) and deep genome sequencing have allowed for the detection of viral genomes in water samples (1, 4, 9, 10).Enteric viruses include rotavirus, adenovirus, coxsackievirus, and norovirus (4). Rotavirus is the leading cause of diarrhea hospitalizations for children under 5 years of age and is responsible for the deaths of an estimated 453,000 children each year (11). Despite this impact on human health, little is known as to what conditions result in the best disinfection of rotavirus in wastewater and drinking water. In addition, it is unclear how a disinfectant functions on a molecular level to inactivate rotavirus.Rotavirus is a nonenveloped virus with a double-stranded, segmented RNA genome. Six structural proteins make up the capsid, including VP4 and VP7. For rotavirus to attach to its host cell, VP4 binds to the host cell receptors, and after a multistep process involving the VP4, VP7, and other cellular proteins, rotavirus enters the host cell through endocytosis. Upon penetration, rotavirus loses its outer capsid (VP7 and VP4), and the double-layered particle is transported to the cell cytopl...

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“…It is thought that the initial contact of RRV with the cell surface is through an SA-containing cell receptor using the VP8 domain of VP4. Gangliosides have been suggested to play this role (43,44), although recent data suggest that gangliosides could function during rotavirus entry at a postattachment step (45). The initial interaction of VP8 with SA is proposed to induce a subtle conformational change on VP4 that allows the virus to interact subsequently with integrin ␣2␤1 through the DGE motif on VP5 (42).…”

Section: Discussionmentioning

confidence: 99%

The Spike Protein VP4 Defines the Endocytic Pathway Used by Rotavirus To Enter MA104 Cells

Díaz-Salinas

Romero

Espinosa

et al. 2013

J Virol

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bRotaviruses are internalized into MA104 cells by endocytosis, with different endocytic pathways used depending on the virus strain. The bovine rotavirus UK strain enters cells through a clathrin-mediated endocytic process, while the simian rhesus rotavirus (RRV) strain uses a poorly defined endocytic pathway that is clathrin and caveolin independent. The viral surface protein VP7 and the spike protein VP4 interact with cellular receptors during cell binding and penetration. To determine the viral protein that defines the mechanism of internalization, we used a panel of UK ؋ RRV reassortant viruses having different combinations of the viral structural proteins. Characterization of the infectivities of these reassortants in MA104 cells either transfected with a small interfering RNA (siRNA) against the heavy chain of clathrin or incubated with hypertonic medium that destabilizes the clathrin coat clearly showed that VP4 determines the pathway of virus entry. Of interest, the characterization of Nar3, a sialic acid-independent variant of RRV, showed that a single amino acid change in VP4 shifts the route of entry from being clathrin dependent to clathrin independent. Furthermore, characterizations of several additional rotavirus strains that differ in their use of cellular receptors showed that all entered cells by clathrin-mediated endocytosis, suggesting that diverse VP4-cell surface interactions can lead to rotavirus cell entry through this endocytic pathway.

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Gangliosides Have a Functional Role during Rotavirus Cell Entry

Cited by 63 publications

References 45 publications

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

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

Solar and Temperature Treatments Affect the Ability of Human Rotavirus Wa To Bind to Host Cells and Synthesize Viral RNA

The Spike Protein VP4 Defines the Endocytic Pathway Used by Rotavirus To Enter MA104 Cells

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