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

Makoto Hijikata, Hiroto Mizushima, Yasunori Tanji, Yasumasa Komoda, Yuji Hirowatari, Tsuyoshi Akagi, Nobuyuki Kato, Koichi Kimura, Kunitada Shimotohno

Research output: Contribution to journalArticle

301 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)10773-10777
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number22
Publication statusPublished - Nov 15 1993
Externally publishedYes

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Protein C
Hepacivirus
Polyproteins
Membranes
Peptides
Protein Precursors
Serine Proteases
Viral Proteins
Immunoprecipitation
Cultured Cells
Proteins
Plasmids
Peptide Hydrolases
Antibodies
In Vitro Techniques

Keywords

  • RNA virus
  • Serine proteinase

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Hijikata, M., Mizushima, H., Tanji, Y., Komoda, Y., Hirowatari, Y., Akagi, T., ... Shimotohno, K. (1993). 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, 90(22), 10773-10777.

Proteolytic processing and membrane association of putative nonstructural proteins of hepatitis C virus. / Hijikata, Makoto; Mizushima, Hiroto; Tanji, Yasunori; Komoda, Yasumasa; Hirowatari, Yuji; Akagi, Tsuyoshi; Kato, Nobuyuki; Kimura, Koichi; Shimotohno, Kunitada.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 22, 15.11.1993, p. 10773-10777.

Research output: Contribution to journalArticle

Hijikata, M, Mizushima, H, Tanji, Y, Komoda, Y, Hirowatari, Y, Akagi, T, Kato, N, Kimura, K & Shimotohno, K 1993, '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, pp. 10773-10777.
Hijikata, Makoto ; Mizushima, Hiroto ; Tanji, Yasunori ; Komoda, Yasumasa ; Hirowatari, Yuji ; Akagi, Tsuyoshi ; Kato, Nobuyuki ; Kimura, Koichi ; Shimotohno, Kunitada. / Proteolytic processing and membrane association of putative nonstructural proteins of hepatitis C virus. In: Proceedings of the National Academy of Sciences of the United States of America. 1993 ; Vol. 90, No. 22. pp. 10773-10777.
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