Gene mapping of the putative structural region of the hepatitis C virus genome by in vitro processing analysis

Makoto Hijikata, Nobuyuki Kato, Yuko Ootsuyama, Masanori Nakagawa, Kunitada Shimotohno

Research output: Contribution to journalArticle

566 Citations (Scopus)

Abstract

Processing of the putative structural proteins of hepatitis C virus was examined by using an in vitro expression system. An RNA transcript for cell-free translation was prepared from a cDNA construct that encompasses the region encoding the 980 amino-terminal residues of the viral polyprotein precursor. Processing of the in vitro translation product proceeded cotranslationally in the presence of microsomal membranes and generated four major membrane-associated products. Two of these four major products, named gp35 and gp70, were shown to be transported into microsomes and heavily glycosylated, suggesting that the processing events are partly mediated by the signal peptidase of the endoplasmic reticulum. The other two products, p19 and p21, were probably associated with the outer surface of the microsomal membrane. Analysis of processed proteins translated from a series of truncated forms of the cDNA construct as well as determination of amino-terminal amino acid sequences of gp35 and gp70 indicated that these four products are arranged from the amino-terminal end of the polyprotein precursor in the order: NH2-p22-gp35-gp70-p19. Both gp35 and gp70 could be candidates of initially processed forms of envelope proteins of the hepatitis C virus.

Original languageEnglish
Pages (from-to)5547-5551
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number13
DOIs
Publication statusPublished - Jul 1 1991
Externally publishedYes

Keywords

  • Proteins glycoproteins
  • RNA
  • Signal peptidase
  • Virus structural

ASJC Scopus subject areas

  • General

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