title: Substitution at aspartic acid 1128 in the SARS coronavirus spike glycoprotein mediates escape from a S2 domain-targeting neutralizing monoclonal antibody.
abstract: The Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) is the etiological agent for the infectious disease, SARS, which first emerged 10 years ago. SARS-CoV is a zoonotic virus that has crossed the species barriers to infect humans. Bats, which harbour a diverse pool of SARS-like CoVs (SL-CoVs), are believed to be the natural reservoir. The SARS-CoV surface Spike (S) protein is a major antigenic determinant in eliciting neutralizing antibody production during SARS-CoV infection. In our previous work, we showed that a panel of murine monoclonal antibodies (mAbs) that target the S2 subunit of the S protein are capable of neutralizing SARS-CoV infection in vitro (Lip KM et al, J Virol. 2006 Jan; 80(2): 941-50). In this study, we report our findings on the characterization of one of these mAbs, known as 1A9, which binds to the S protein at a novel epitope within the S2 subunit at amino acids 1111-1130. MAb 1A9 is a broadly neutralizing mAb that prevents viral entry mediated by the S proteins of human and civet SARS-CoVs as well as bat SL-CoVs. By generating mutant SARS-CoV that escapes the neutralization by mAb 1A9, the residue D1128 in S was found to be crucial for its interaction with mAb 1A9. S protein containing the substitution of D1128 with alanine (D1128A) exhibited a significant decrease in binding capability to mAb 1A9 compared to wild-type S protein. By using a pseudotyped viral entry assay, it was shown that the D1128A substitution in the escape virus allows it to overcome the viral entry blockage by mAb 1A9. In addition, the D1128A mutation was found to exert no effects on the S protein cell surface expression and incorporation into virion particles, suggesting that the escape virus retains the same viral entry property as the wild-type virus.
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University Health System (NUHS), National University of Singapore, Singapore.
authors.forename: Oi-Wing
authors.initials: OW
authors.lastname: Ng
authors.affiliation: Institute of Molecular and Cell Biology, A*STAR (Agency for Science, Technology and Research), Singapore.
authors.forename: Choong-Tat
authors.initials: CT
authors.lastname: Keng
authors.affiliation: Centre of Influenza Research, School of Public Health, The University of Hong Kong, Hong Kong, China.
authors.forename: Cynthia Sau-Wai
authors.initials: CS
authors.lastname: Leung
authors.affiliation: Centre of Influenza Research, School of Public Health, The University of Hong Kong, Hong Kong, China.
authors.forename: J S Malik
authors.initials: JS
authors.lastname: Peiris
authors.affiliation: Centre of Influenza Research, School of Public Health, The University of Hong Kong, Hong Kong, China.
authors.forename: Leo Lit Man
authors.initials: LL
authors.lastname: Poon
authors.affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, National University Health System (NUHS), National University of Singapore, Singapore; Institute of Molecular and Cell Biology, A*STAR (Agency for Science, Technology and Research), Singapore.
authors.forename: Yee-Joo
authors.initials: YJ
authors.lastname: Tan
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology; D057134:Antibodies, Neutralizing / Q000502:physiology*; D000914:Antibodies, Viral / Q000502:physiology; D001224:Aspartic Acid / Q000737:chemistry*; D016466:CHO Cells; D002685:Chiroptera / Q000821:virology; D002522:Chlorocebus aethiops; D003412:Cricetulus; D018604:Epitope Mapping; D057809:HEK293 Cells; D006801:Humans; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D016415:Sequence Alignment; D064370:Spike Glycoprotein, Coronavirus / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D014709:Vero Cells; D014771:Virion / Q000235:genetics / Q000472:pathogenicity; D045949:Viverridae / Q000821:virology
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D064370:Spike Glycoprotein, Coronavirus; D001224:Aspartic Acid