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PMID: 2688301 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Identification of essential residues in potyvirus proteinase HC-Pro by site-directed mutagenesis.

Virology ·Vol. 173 ·No. 2 ·1989-12-00 ·Pages 692-9

Oh CS, Carrington JC

Abstract

Two virus-encoded proteinases are responsible for proteolysis of potyvirus polyproteins. One of these, HC-Pro, is a multifunctional protein that autolytically cleaves at its carboxyl-terminus (J.C. Carrington et al., 1989, EMBO J. 8, 365-370). To identify the class of proteinase to which HC-Pro belongs, tobacco etch virus (TEV) HC-Pro mutants containing single amino acid substitutions at serine, cysteine, aspartic acid, and histidine positions were synthesized by in vitro transcription and translation and were tested for autoproteolytic activity. Combinations of these residues are constituents of the active sites of diverse groups of cellular and viral proteinases. Only those positions that were strictly conserved among four potyvirus HC-Pro proteolytic domains (for which sequences have been deduced) were mutagenized. Of the 19 mutant proteinases synthesized and tested, only those with alterations at Cys-649 and His-722 were defective for HC-Pro autolytic activity. Most of the other mutant proteinases exhibited no impairments in processing kinetics experiments. The spectrum of essential residues, as defined by this genetic analysis, supports the hypothesis that HC-Pro most closely resembles members of the cysteine-type family of proteinases.

MeSH Terms
Amino Acid Sequence Base Sequence DNA Mutational Analysis DNA, Viral/genetics Endopeptidases/genetics,metabolism Molecular Sequence Data Plant Viruses/enzymology,genetics Protein Biosynthesis Transcription, Genetic
Chemicals
DNA, Viral Endopeptidases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Oh C S
Department of Biology, Texas A&M University, College Station 77843.
Carrington J C
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Article Info
Journal
Virology
Abbr.
Virology
ISSN
0042-6822
DOI
10.1016/0042-6822(89)90582-5
Published
1989-12-00
Pages
692-9
Language
English
Region
United States
NLM ID
0110674
PMCID
PMC7131030
Subset
IM
Grants
NIAID NIH HHS · AI27842 · United States
External Links
PubMed source DOI: 10.1016/0042-6822(89)90582-5 PMC Full Text View journal details
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