title: Relative roles of GM1 ganglioside, N-acylneuraminic acids, and α2β1 integrin in mediating rotavirus infection.
abstract: N-acetyl- and N-glycolylneuraminic acids (Sia) and α2β1 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 α-d-N-acetylneuraminide treatment, but did not alter α2β1 usage. CTB reduced the infectivity of these viruses. Aceramido-GM1 inhibited Wa and RV-3 infectivity in untreated and sialidase-treated cells, and GM1 supplementation increased their infectivity, demonstrating the importance of GM1 for infection. Wa recognition of α2β1 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.
  
  
  OBJECTIVE
  Rotaviruses, 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.
authors: Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity at The University of Melbourne, Melbourne, Victoria, Australia.
authors: Fiona E
authors: FE
authors: Fleming
authors: Raphael
authors: R
authors: Böhm
authors: Vi T
authors: VT
authors: Dang
authors: Gavan
authors: G
authors: Holloway
authors: Thomas
authors: T
authors: Haselhorst
authors: Paul D
authors: PD
authors: Madge
authors: Jaigeeth
authors: J
authors: Deveryshetty
authors: Xing
authors: X
authors: Yu
authors: Helen
authors: H
authors: Blanchard
authors: Mark
authors: M
authors: von Itzstein
authors: Barbara S
authors: BS
authors: Coulson
mesh_terms: D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D005732:Gangliosides / Q000378:metabolism*; D006801:Humans; D038982:Integrin alpha2beta1 / Q000235:genetics / Q000378:metabolism*; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D009438:Neuraminic Acids / Q000378:metabolism*; D011485:Protein Binding; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism*; D012401:Rotavirus / Q000235:genetics / Q000502:physiology*; D012400:Rotavirus Infections / Q000235:genetics / Q000378:metabolism* / Q000821:virology
chemical_list: D036022:Capsid Proteins; D005732:Gangliosides; D038982:Integrin alpha2beta1; D009438:Neuraminic Acids; D011991:Receptors, Virus; C060339:VP4 protein, Rotavirus; D019158:N-Acetylneuraminic Acid