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PMID: 7504283 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Proteolytic processing and membrane association of putative nonstructural proteins of hepatitis C virus.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 90 ·No. 22 ·1993-11-15 ·Pages 10773-7

Hijikata M, Mizushima H, Tanji Y, Komoda Y, Hirowatari Y, Akagi T, Kato N, Kimura K, Shimotohno K

Abstract

By using a plasmid-based transient protein expression system in cultured cells and an in vitro transcription/translation system, we analyzed the proteolytic processing of the putative nonstructural protein region of the precursor polyprotein from a Japanese type of hepatitis C virus. In addition to the previously reported viral proteins, p21 and p70, we identified products of 4 kDa (p4), 27 kDa (p27), 56 kDa (p56), 58 kDa (p58), and 66 kDa (p66). These products were produced in a viral serine proteinase (proteinase 2)-dependent manner from the region downstream of p70 in the precursor polyprotein and were arranged as NH2-p70-p4-p27-p58(p56)-p66-COOH as determined with region-specific antibodies. We showed that p56 was an N-terminally truncated form of p58, which suggested that a small polypeptide of 2 kDa (p2) was produced from the N-terminal part of p58. Cleavage between p4 and p27 was inefficient in vitro and we saw the 31-kDa precursor polypeptide (p31) accumulate. Furthermore, efficient cleavage at this site in vivo required the presence of p58/p56. Immunoprecipitation analysis in vitro also suggested the mutual interaction of those nonstructural protein products. An especially close association of p4 with p70 may contribute to association of p70 with microsomal membranes.

MeSH Terms
Animals Base Sequence Cell Line Cell Membrane/metabolism Chlorocebus aethiops Cloning, Molecular DNA Primers/chemistry Epitopes Hepacivirus/metabolism In Vitro Techniques Membrane Proteins/metabolism Molecular Sequence Data Protein Processing, Post-Translational Sequence Deletion Viral Nonstructural Proteins/metabolism
Chemicals
DNA Primers Epitopes Membrane Proteins Viral Nonstructural Proteins
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Hijikata M
Virology Division, National Cancer Center Research Institute, Tokyo, Japan.
Mizushima H
Tanji Y
Komoda Y
Hirowatari Y
Akagi T
Kato N
Kimura K
Shimotohno K
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.90.22.10773
Published
1993-11-15
Pages
10773-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC47860
Subset
IM
Databases
GENBANK
D16435
External Links
PubMed source DOI: 10.1073/pnas.90.22.10773 PMC Full Text View journal details
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