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. 2002;8(2):143–149. doi: 10.1080/13550280290049598

Further in vitro characterization of mouse hepatitis virus papain-like proteinase 1: Cleavage sequence requirements within PP1a

Henry Teng 1, Susan R Weiss 1,✉
PMCID: PMC7095356  PMID: 11935466

Abstract

Proteolytic processing of the mouse hepatitis virus strain A59 (MHV-A59) replicase gene product, pp1a, results in polypeptides p28, p65, p50, and p240 in infected cells. Based on previously identified p28 and p65 cleavage sites, a p50 cleavage site was proposed to occur between Ala-1262 and Ala-1263. Results of mutagenesis and in vitro cleavage assays show that PLP-1 was able to cleave in trans when the proposed p50 cleavage sequence replaced the p28 cleavage sequence. Mutagenesis was also used to investigate cleavage between Gly-904 and Val-905, a cleavage site predicted to produce a precursor of p65, p72, that was detected in cells infected with MHV strain JHM, but not with MHV-A59. No cleavage could be detected using substrate that carried both the p65 site and the predicted p72 cleavage sequence. Thus, it appeared that PLP-1 could recognize the proposed p50 sequence but not the predicted p72 site under the in vitro conditions used.

Keywords: proteolytic processing, viral papain-like proteinase

Footnotes

This work was supported by NIH grant AI-17418.

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