a single amino acid mutation in the spike protein of coronavirus infectious bronchitis virus hampers its maturation and incorporation into virions at the nonpermissive temperature：一个单一的氨基酸变异冠状病毒传染性支气管炎病毒s蛋白阻碍其成熟和掺入病毒粒子在nonpermissive温度.pdfVIP

Virology 326 (2004) 288–298 /locate/yviro A single amino acid mutation in the spike protein of coronavirus infectious bronchitis virus hampers its maturation and incorporation into virions at the nonpermissive temperature S. Shen, Y.C. Law, D.X. Liu * Institute of Molecular and Cell Biology, 117609, Singapore Received 10 March 2004; returned to author for revision 5 April 2004; accepted 3 June 2004 Available online 14 July 2004 Abstract The spike (S) glycoprotein of coronavirus is responsible for receptor binding and membrane fusion. A number of variants with deletions and mutations in the S protein have been isolated from naturally and persistently infected animals and tissue cultures. Here, we report the emergence and isolation of two temperature sensitive (ts) mutants and a revertant in the process of cold-adaptation of coronavirus infectious bronchitis virus (IBV) to a monkey kidney cell line. The complete sequences of wild type (wt) virus, two ts mutants, and the revertant were compared and variations linked to phenotypes were mapped. A single amino acid reversion (L294-to-Q) in the S protein is sufficient to abrogate the ts phenotype. Interestingly, unlike wt virus, the revertant grows well at and below 32 jC, the permissive temperature, as it carries other mutations in multiple genes that might be associated with the cold-adaptation phenotype. If the two ts mutants were allowed to enter cells at 32 jC, the S protein was synthesized, core-glycosylated and at least partially modified at 40 jC. H