Hepatitis C virus-encoded nonstructural protein NS4A has versatile functions in viral protein processing

Y. Tanji, M. Hijikata, Shinya Satoh, T. Kaneko, K. Shimotohno

Research output: Contribution to journalArticle

216 Citations (Scopus)

Abstract

A transient protein expression system in COS-1 cells was used to study the role of hepatitis C virus (HCV)-encoded NS4A protein on HCV nonstructural polyprotein processing. By analyzing the protein expression and processing of a deletion mutant polypeptide, NSΔ4A, which encodes the entire putative HCV nonstructural polyprotein except the region encoding NS4A, the versatile functions of NS4A were revealed. Most of the NS3 processed from NSΔ4A was localized in the cytosol fraction and was degraded promptly. Coproduction of NS4A stabilizes NS3 and assists in its localization in the membrane. NS4A was found to be indispensable for cleavage at the 4B/5A site but not essential for cleavage at the 5A/5B site in NSΔ4A. The functioning of NS4A as a cofactor for cleavage at the 4B/5A site was also observed when 30 amino acids around this site was used as a substrate and a serine proteinase domain of 167 amino acids, from Gly-1049 to Ser-1215, was used as an enzyme protein, suggesting that possible domains for the interaction of NS4A were in those regions of the enzyme protein (NS3) and/or the substrate protein. Two proteins, p58 and p56, were produced from NS5A. For the production of p58, equal or excess molar amounts of NS4A relative to NSΔ4A were required. Deletion analysis of NS4A revealed a minimum functional domain of NS4A of 10 amino acids, from Gly-1678 to Ile-1687

Original languageEnglish
Pages (from-to)1575-1581
Number of pages7
JournalJournal of Virology
Volume69
Issue number3
Publication statusPublished - 1995
Externally publishedYes

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Hepatitis C virus
viral proteins
Viral Proteins
Hepacivirus
Proteins
proteins
Polyproteins
amino acids
protein synthesis
Amino Acids
serine proteinases
enzymes
cytosol
polypeptides
COS Cells
Serine Proteases
Enzymes
Cytosol
mutants
Peptides

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Hepatitis C virus-encoded nonstructural protein NS4A has versatile functions in viral protein processing. / Tanji, Y.; Hijikata, M.; Satoh, Shinya; Kaneko, T.; Shimotohno, K.

In: Journal of Virology, Vol. 69, No. 3, 1995, p. 1575-1581.

Research output: Contribution to journalArticle

Tanji, Y, Hijikata, M, Satoh, S, Kaneko, T & Shimotohno, K 1995, 'Hepatitis C virus-encoded nonstructural protein NS4A has versatile functions in viral protein processing', Journal of Virology, vol. 69, no. 3, pp. 1575-1581.
Tanji, Y. ; Hijikata, M. ; Satoh, Shinya ; Kaneko, T. ; Shimotohno, K. / Hepatitis C virus-encoded nonstructural protein NS4A has versatile functions in viral protein processing. In: Journal of Virology. 1995 ; Vol. 69, No. 3. pp. 1575-1581.
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