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PMID: 8386278 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Characterization of the hepatitis C virus-encoded serine proteinase: determination of proteinase-dependent polyprotein cleavage sites.

Journal of virology ·Vol. 67 ·No. 5 ·1993-05-00 ·Pages 2832-43

Grakoui A, McCourt DW, Wychowski C, Feinstone SM, Rice CM

Abstract

Processing of the hepatitis C virus (HCV) H strain polyprotein yields at least nine distinct cleavage products: NH2-C-E1-E2-NS2-NS3-NS4A-NS4B-NS5A-NS5B-CO OH. As described in this report, site-directed mutagenesis and transient expression analyses were used to study the role of a putative serine proteinase domain, located in the N-terminal one-third of the NS3 protein, in proteolytic processing of HCV polyproteins. All four cleavages which occur C terminal to the proteinase domain (3/4A, 4A/4B, 4B/5A, and 5A/5B) were abolished by substitution of alanine for either of two predicted residues (His-1083 and Ser-1165) in the proteinase catalytic triad. However, such substitutions have no observable effect on cleavages in the structural region or at the 2/3 site. Deletion analyses suggest that the structural and NS2 regions of the polyprotein are not required for the HCV NS3 proteinase activity. NS3 proteinase-dependent cleavage sites were localized by N-terminal sequence analysis of NS4A, NS4B, NS5A, and NS5B. Sequence comparison of the residues flanking these cleavage sites for all sequenced HCV strains reveals conserved residues which may play a role in determining HCV NS3 proteinase substrate specificity. These features include an acidic residue (Asp or Glu) at the P6 position, a Cys or Thr residue at the P1 position, and a Ser or Ala residue at the P1' position.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cells, Cultured Hepacivirus/enzymology,genetics Humans Molecular Sequence Data Mutagenesis, Site-Directed Protein Precursors/metabolism Protein Processing, Post-Translational RNA Helicases Recombinant Proteins/metabolism Sequence Analysis Sequence Homology, Amino Acid Serine Endopeptidases Vaccinia virus/genetics Viral Nonstructural Proteins/genetics,metabolism
Chemicals
NS2B protein, flavivirus NS3 protein, flavivirus Protein Precursors Recombinant Proteins Viral Nonstructural Proteins Serine Endopeptidases RNA Helicases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Grakoui A
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110-1093.
McCourt D W
Wychowski C
Feinstone S M
Rice C M
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.67.5.2832-2843.1993
Published
1993-05-00
Pages
2832-43
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237608
Subset
IM
Grants
NCI NIH HHS · CA57973 · United States
External Links
PubMed source DOI: 10.1128/JVI.67.5.2832-2843.1993 PMC Full Text View journal details
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