PMID: 8985369Jan 1, 1997Paper

Analysis of cleavage site mutations between the NC and PR Gag domains of Rous sarcoma virus

Journal of Virology
G SchatzV M Vogt

Abstract

In retroviruses, the viral protease (PR) is released as a mature protein by cleavage of Gag, Gag-Pro, or Gag-Pro-Pol precursor polypeptides. In avian sarcoma and leukemia viruses (ASLV), PR forms the C-terminal domain of Gag. Based on the properties of a mutation (cs22) in the cleavage site between the upstream NC domain and the PR domain, the proteolytic liberation of PR previously was inferred to be essential for processing of Gag and Pol proteins. To study this process in more detail, we have analyzed the effects that several mutations at the NC-PR cleavage site have on proteolytic processing in virus-like particles expressed in COS and quail cells. Mutant Gag proteins carrying the same mutations also were synthesized in vitro and tested for processing with purified PR. In both types of studies, N-terminal sequencing of the liberated PR domain was carried out to exactly identify the site of cleavage. Finally, synthetic peptides corresponding to the mutant proteins were assessed for the ability to act as substrates for PR. The results were all consistent and led to the following conclusions. (i) In vivo, if normal processing between NC and PR is prevented by mutations, limited cleavage occurs at a previously unrecognized alte...Continue Reading

References

Oct 1, 1990·Journal of Virology·L StewartV M Vogt
Apr 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·H G Kräusslich
Jan 1, 1990·Annual Review of Biophysics and Biophysical Chemistry·D R Davies
Aug 31, 1990·Biochemical and Biophysical Research Communications·L H PhylipA V Broadhurst
Jun 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·M KotlerA M Skalka
Aug 1, 1988·Journal of Virology·M KotlerA M Skalka
Aug 16, 1994·Proceedings of the National Academy of Sciences of the United States of America·J M LouisD M Jerina
Jan 1, 1993·Annual Review of Biochemistry·A Wlodawer, J W Erickson

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