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An Amino Acid Substitution in the Coding Region of the E2 Glycoprotein Adapts Ross River Virus To Utilize Heparan Sulfate as an Attachment Moiety

Heil, Marintha L. and Albee, Alison and Strauss, James H. and Kuhn, Richard J. (2001) An Amino Acid Substitution in the Coding Region of the E2 Glycoprotein Adapts Ross River Virus To Utilize Heparan Sulfate as an Attachment Moiety. Journal of Virology, 75 (14). pp. 6303-6309. ISSN 0022-538X. PMCID PMC114352.

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Passage of Ross River virus strain NB5092 in avian cells has been previously shown to select for virus variants that have enhanced replication in these cells. Sequencing of these variants identified two independent sites that might be responsible for the phenotype. We now demonstrate, using a molecular cDNA clone of the wild-type T48 strain, that an amino acid substitution at residue 218 in the E2 glycoprotein can account for the phenotype. Substitutions that replaced the wild-type asparagine with basic residues had enhanced replication in avian cells while acidic or neutral residues had little or no observable effect. Ross River virus mutants that had increased replication in avian cells also grew better in BHK cells than the wild-type virus, whereas the remaining mutants were unaffected in growth. Replication in both BHK and avian cells of Ross River virus mutants N218K and N218R was inhibited by the presence of heparin or by the pretreatment of the cells with heparinase. Binding of the mutants, but not of the wild type, to a heparin-Sepharose column produced binding comparable to that of Sindbis virus, which has previously been shown to bind heparin. Replication of these mutants was also adversely affected when they were grown in a CHO cell line that was deficient in heparan sulfate production. These results demonstrate that amino acid 218 of the E2 glycoprotein can be modified to create an heparan sulfate binding site and this modification expands the host range of Ross River virus in cultured cells to cells of avian origin.

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Additional Information:Copyright © 2001, American Society for Microbiology. Received 26 January 2001/Accepted 19 April 2001 We thank Andrew P. Byrnes and Diane E. Griffin for the generous gift of the CHO-K1 cells and the mutant cell line CHO18.4m. This research was supported by Public Health Service grants AI 20612 to J.H.S. and GM 56279 to R.J.K.
Funding AgencyGrant Number
NIHAI 20612
NIHGM 56279
Issue or Number:14
PubMed Central ID:PMC114352
Record Number:CaltechAUTHORS:HEIjvir01
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:2192
Deposited By: Archive Administrator
Deposited On:14 Mar 2006
Last Modified:06 Jul 2022 21:19

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