Identification of a New Region of SARS-CoV S Protein Critical for Viral Entry

JOURNAL OF MOLECULAR BIOLOGY

卷:394

Article

Guo, Ying²  Tisoncik, Jennifer²  McReynolds, Susanna¹  Farzan, Michael³  Prabhakar, Bellur S.²  Gallagher, Thomas⁴  Rong, Lijun²  Caffrey, Michael¹ 

[1] Univ Illinois, Coll Med, Dept Biochem & Mol Genet, Chicago, IL 60607 USA

[2] Univ Illinois, Coll Med, Dept Microbiol & Immunol, Chicago, IL 60607 USA

[3] Partners AIDS Res Ctr, Cambridge, MA 02139 USA

[4] Loyola Univ, Med Ctr, Dept Microbiol & Immunol, Maywood, IL 60153 USA

关键词: viral entry;    SARS-CoV;    envelope;    mutagenesis;    spike;   

DOI  :  10.1016/j.jmb.2009.10.032

来源: Elsevier

【 摘 要 】

Infection by severe acute respiratory syndrome coronavirus (SARS-CoV) is initiated by specific interactions between the SARS-CoV spike (S) protein and its receptor ACE2. In this report, we screened a peptide library representing the SARS-CoV S protein sequence using a human immunodeficiency virus-based pseudotyping system to identify specific regions that affect viral entry. One of the 169 peptides screened, peptide 9626 (S residues 217-234), inhibited SARS-CoV S-mediated entry of the pseudotyped virions in 293T cells expressing a functional SARS-CoV receptor (human angio tensin-converting enzyme 2) in a dose-dependent manner (IC50 similar to 11 mu M). Alanine scanning mutagenesis was performed to assess the roles of individual residues within this region of S, which was previously uncharacterized. The effects included significant reductions in expression (K223A), viral incorporation (L218A, I230A, and N232A), and reduced viral entry (L224A, L226A, I228A, T231A, and F233A). Taken together, these results reveal a new region of the S protein that is crucial for SARS-CoV entry. (C) 2009 Elsevier Ltd. All rights reserved.

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