Identification of the host lipid raft protein which interacts with hepatitis C virus protein

2010 Fiscal Year Final Research Report

• Project/Area Number: 20590767
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: University of Yamanashi
• Principal Investigator: INOUE Taisuke University of Yamanashi, 大学院・医学工学総合研究部, 講師 (90422691)
• Co-Investigator(Kenkyū-buntansha): ENOMOTO Nobuyuki 山梨大学, 大学院・医学工学総合研究部, 教授 (20251530)
• Project Period (FY): 2008 – 2010
• Keywords: C型肝炎ウイルス / HCV NS5B / α-tubulin / 複製複合体 / プロテオミクス解析

Research Abstract

Hepatitis C virus (HCV) non structural proteins and host proteins are believed to be involved in HCV replication complex on lipid raft. To identify these host proteins that interact with HCV proteins, we employed the proteomics approach, combination of tandem affinity purification (TAP) method and mass spectrometry analysis. α-tubulin was identified as a novel HCV NS5B binding protein. Interaction between these two proteins was confirmed by the immunohistchemistry study and the co-immunoprecipitation study. To examine the effect of the interaction between these two proteins, α-tubulin expression was suppressed by siRNA on the HCV replicon stably expressing cells. By real-time PCR method, the HCV replicon RNA level was suppressed to 60% on the α-tubulin knock down cells. These results suggest that α-tubulin interacts with HCV NS5B protein and this interaction may be important for the HCV RNA replication. Further studies to identify direct interacting domain are required.

Research Products

• [Journal Article] Analysis of the complete open reading frame of hepatitis C virus in genotype 2a infection reveals critical sites influencing the response to peginterferon and ribavirin therapy. (2011)
  • Author(s): Kadokura M, Maekawa S, Sueki R, Miura M, Komase K, Shindo H, Amemiya F, Uetake T, Inoue T, Sakamoto M, Nakagawa M, Sakamoto N, Watanabe M, Enomoto N.
  • Journal Title: Hepatol Int. Mar
  • Peer Reviewed
• [Journal Article] DICKKOPF-4 and -2 genes are upregulated in human colorectal cancer. (2009)
  • Author(s): Matsui A, Yamaguchi T, Markawa S, Miyazaki C, Takano S, Uetake T, Inoue T, Otaka M, Otsuka H, Sato T, Yamashita A, Takahashi Y, Enomoto N.
  • Journal Title: Cancer Sci. 100(10) Pages: 1923-1930
  • Peer Reviewed
• [Journal Article] Targeting lipid metabolism in the treatment of hepatitis C virus infection. (2008)
  • Author(s): Amemiya F, Maekawa S, Itakura Y, Kanayama A, Matsui A, Takano S, Yamaguchi T, Itakura J, Kitamura T, Inoue T, Sakamoto M, Yamauchi K, Okada S, Yamashita A, Sakamoto N, Itoh M, Enomoto N.
  • Journal Title: J Infect Dis. 197(3) Pages: 631-370
  • Peer Reviewed
• [Presentation] α-tubulinn is Identified as an Interacting Partner of Hepatitis C NS5B by Protemics Analysis. (2007)
  • Author(s): Inoue T, Amemiya F, Stanley M. Lemon, Makino S, Enomoto N.
  • Organizer: 17th APASL Conference
  • Place of Presentation: Kyoto, Japan.
  • Year and Date: 20070327-20070330
• [Presentation] Proteomics analysis identifies an interaction between Hepatitis C Virus NS5B and alfatubulin. (2006)
  • Author(s): Inoue T, Huang C, Sugiyama K, Mizutani T, Soon B Hwang, Enomoto N, Makino S.
  • Organizer: 13th International Meeting on Hepatitis C virus and Related Viruses
  • Place of Presentation: Cairns, Australia.
  • Year and Date: 20060827-20060831
• [Book] Proteomics解析の手法を用いたC型肝炎ウイルス蛋白と宿主細胞蛋白の相互作用の検討.(消化器発癌における炎症・再生・細胞応答の役割小俣政男アークメディア) (2007)
  • Author(s): 井上泰輔、Cheng Huang, Kazuo Sugiyama, Tetsuya Mizutani, Soon B Hwang, Stanly M Lemon, Shinji Makino, 榎本信幸
  • Publisher: 学術アドバイザー