Efficient infection of buffalo rat liver-resistant cells by encephalomyocarditis virus requires binding to cell surface sialic acids.
In contrast to the production of virus and cell lysis seen in baby hamster kidney cells (BHK-21) infected with the strain 1086C of encephalomyocarditis virus (EMCV), in buffalo rat liver cells (BRL) neither virus replication nor cytopathic effects were observed. After 29 passages in BRL cells, each alternating with boosts of the recovered virus in BHK-21 cells, the virus acquired the ability to replicate effectively in BRL cells, attaining virus titres comparable to those in BHK-21 cells and producing complete cell destruction. The binding of virus on BRL cells was increased after adaptation and was similar to that observed on BHK-21 cells. Treatment of BRL cells with sialidase resulted in an 87 % reduction in virus binding and inhibition of infection. Sequence analyses revealed three mutations in the VP1 amino acid sequence of the adapted virus at positions 49 (Lys-->Glu), 142 (Leu-->Phe) and 180 (Ile-->Ala). The residue 49 is exposed at the surface of the capsid and is known to be part of a neutralization epitope. These results suggest that the adaptation of EMCV to BRL cells may have occurred through a mutation in a neutralizing site that confers to the virus a capacity to interact with cell surface sialic acid residues. Taken together, these data suggest a link between virus neutralization site, receptor binding and cell permissivity to infection.
Guy
Monique
M
UMR 1161 INRA, AFSSA, ENVA, Ecole Nationale Vétérinaire, 7 Avenue Général de Gaulle, 94704 Maisons-Alfort Cedex, France.
Chilmonczyk
Stefan
S
Crucière
Catherine
C
Eloit
Marc
M
Bakkali-Kassimi
Labib
L
D000220:Adaptation, Biological; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D036022:Capsid Proteins / Q000235:genetics; D002460:Cell Line; D006224:Cricetinae; D003588:Cytopathogenic Effect, Viral; D004252:DNA Mutational Analysis; D004686:Encephalomyocarditis virus / Q000502:physiology*; D018985:Epitopes, B-Lymphocyte / Q000235:genetics; D022781:Hepatocytes / Q000821:virology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D009500:Neutralization Tests; D017434:Protein Structure, Tertiary; D051381:Rats; D011915:Rats, Inbred BUF; D010948:Viral Plaque Assay; D053585:Virus Attachment
D036022:Capsid Proteins; D018985:Epitopes, B-Lymphocyte; C096256:VP1 protein, encephalomyocarditis virus; D019158:N-Acetylneuraminic Acid