Dissect the role of hepatitis C virus NS2 protein in virus assembly

• Project/Area Number: 21590519
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Virology
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: SUZUKI Ryosuke 国立感染症研究所, ウイルス第二部, 主任研究官 (50342902)
• Co-Investigator(Kenkyū-buntansha): SUZUKI Tetsuro 浜松医科大学, 医学部, 教授 (00250184)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2011: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2009: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: HCV / NS2 / 粒子形成 / 感染性粒子 / 宿主因子 / 感染性

Research Abstract

Hepatitis C virus(HCV) nonstructural protein 2(NS2) is a membrane-associated cysteine autoprotease that mediates cleavage of its own carboxyl terminus from NS3.In addition to its role in polyprotein processing, emerging evidence indicates that NS2plays crucial role in assembly of infectious virus particle. However, biological significance of NS2in the HCV assembly remains unclear. In this study, we screened human liver cDNA libraries by split-ubiquitin membrane yeast two-hybrid assay using NS2as a bait, and identified several NS2-interacting proteins. Following siRNA screening identified a membrane-associated cellular protein that is involved in HCV production. Gene silencing of the host factor resulted in no effect on the entry and RNA replication of HCV, suggesting that this host factor is involved in HCV assembly step. Interaction of the host factor with NS2 in mammalian cells was confirmed. Finding of this study may provide important information for understanding the molecular mechanism of assembly and formation of infectious HCV particles.

Research Products

• [Journal Article] Visualization and Measurement of ATP Levels in Living Cells Replicating Hepatitis C Virus Genome RNA (2012)
  • Author(s): 安東友美
  • Journal Title: PLoS Pathogens Volume: 8 Issue: 3 Pages: e1002561-e1002561
  • DOI: 10.1371/journal.ppat.1002561
  • Peer Reviewed
• [Journal Article] Role of the endoplasmic reticulum-associated degradation(ERAD) pathway in degradation of hepatitis C virus envelope proteins and production of virus particles (2011)
  • Author(s): SaeedM, Suzuki R, Watanabe N, Masaki T, Tomonaga M, Muhammad A, Kato T, Matsuura Y, Watanabe H, Wakita T, Suzuki T
  • Journal Title: J Biol Chem Volume: 286 Pages: 37264-73
• [Journal Article] Analyses of mutations selected by passaging a chimeric flavivirus identify mutations that alter infectivity and reveal an interaction between the structural proteins and the nonstructural glycoprotein NS1 (2011)
  • Author(s): Winkelmann ER, Widman DG, Suzuki R, Mason PW
  • Journal Title: Virology Volume: 421(2) Pages: 96-104
• [Journal Article] Analyses of mutations selected by passaging a chimeric flavivirus identify mutations that alter infectivity and reveal an interaction be tween the structural proteins and the nonstructural glycoprotein NS1 (2011)
  • Author(s): Winkelmann ER, Widman DG, Suzuk R, Mason PW
  • Journal Title: Virology Volume: 421 Pages: 96-104
  • Peer Reviewed
• [Journal Article] Role of the ERAD pathway in degradation of hepatitis C virus envelope proteins and production of virus particles (2011)
  • Author(s): Saeed M, Suzuki R, Watanabe N, Masaki T, Tomonaga M, Muhammad A, Kato T, Matsuura Y, Watanabe H, Wakita T, Suzuki T
  • Journal Title: J.Biol.Chem. Volume: 286 Pages: 37264-37273
  • Peer Reviewed
• [Journal Article] Production of infectious hepatitis C virus by using RNA polymerase I-mediated transcription (2010)
  • Author(s): Masaki T, Suzuki R, Saeed M, Mori K, Matsuda M, Aizaki H, Ishii K, Maki N, Miyamura T, Matsuura Y, Wakita T, Suzuki T
  • Journal Title: J Virol Volume: 84 Issue: 11 Pages: 5824-35
  • DOI: 10.1128/jvi.02397-09
• [Journal Article] Involvement of PA28g in the propagation of hepatitis C virus (2010)
  • Author(s): Moriishi K., Shoji I., Mori Y., Suzuki R., Suzuki T., Kataoka C., and Matsuura Y.
  • Journal Title: Hepatology Volume: 52 Issue: 2 Pages: 411-420
  • DOI: 10.1002/hep.23680
  • Peer Reviewed
• [Journal Article] Production of infectious hepatitis C virus by using RNA polymerase I-mediated transcription (2010)
  • Author(s): Masaki T, Suzuki R, Saeed M, Mori K, Matsuda M, Aizaki H, Ishii K, Maki N, Miyamura T, Matsuura Y, Wakita T, Suzuki T
  • Journal Title: J.Virol. 84 Pages: 5824-5835
  • Peer Reviewed
• [Journal Article] Evaluation of hepatitis C virus core antigen assays in detecting recombinant viral antigens of various genotypes (2009)
  • Author(s): Saeed M, Suzuki R, Kondo M, Aizaki H, Kato T, Mizuochi T, Wakita T, Watanabe H, Suzuki T
  • Journal Title: J Clin Microbiol Volume: 47 Issue: 12 Pages: 4141-3
  • DOI: 10.1128/jcm.01437-09
• [Journal Article] Evaluation of hepatitis C virus core antigen assays for detecting recombinant viral antigens derived from various genotypes (2009)
  • Author(s): Saeed M, Suzuki R, Kondo M, Aizaki H, Kato T, Mizuochi T, Wakita T, Suzuki T.
  • Journal Title: J Clin.Micro. 47 Pages: 4141-4143
  • Peer Reviewed
• [Journal Article] Involvement of PA28g in the propagation of hepatitis C virus
  • Author(s): Moriishi K, Shoji I, Mori Y, Suzuki R, Suzuki T, Kataoka C, Matsuura Y.
  • Journal Title: Hepatology (In press)
  • Peer Reviewed
• [Presentation] Identification of lipid droplet? associated membrane proteins that are involved in HCV production (2011)
  • Author(s): Aizaki H, Matsumoto Y, Goto K, Watashi K, Suzuki R, Fukasawa M, Hanada K, Sato S, Takahashi N, Matsuura Y, Motojima K, Miyamura T, Suzuki T, WakitaT
  • Organizer: 18th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle, WA, USA
• [Presentation] Identification and functional analysis of small molecules inhibiting the late step of hepatitis C virus life cycle (2011)
  • Author(s): Watashi K, Uchida N, Suzuki R, Aizaki H, Wakita T
  • Organizer: 18th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle, WA, USA
• [Presentation] Small molecules that elicit strong anti? HCV activity through down? modulation of HCV entry receptor (2011)
  • Author(s): Takebe Y, Uenishi R, Tani H, Suzuki R, Takagi M, Hase S, Liao H, Tsuchiura T, Shinya K, Wakita T, Matsuura Y, Patel A
  • Organizer: 18th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle, WA, USA
• [Presentation] Isolation of a highly infectious hepatitis C virus with adaptive mutations (2011)
  • Author(s): Fukasawa M, Shirasago Y, Saito K, Murakami Y, Fukasawa H, Suzuki T, Suzuki R, Wakita T, Hanada K, Chiba J
  • Organizer: 18th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle, WA, USA
• [Presentation] Identification of novel NS5A? associated proteins in the host? cell membrane fraction and their role in HCV life cycle (2011)
  • Author(s): Goto K, Kimura T, Watashi K, Suzuki R, Yamagoe S, Miyamura T, Moriya K, Yotsuyanagi H, Koike K, Suzuki T, Wakita T, Aizaki H
  • Organizer: 18th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle, WA, USA
• [Presentation] Halopemide inhibited a post? assembly step in hepatitis C virus life cycle (2011)
  • Author(s): Uchida N, Watashi K, Suzuki R, Aizaki H, Chiba J, Wakita T
  • Organizer: 18th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle, WA, USA
• [Presentation] Signal peptidase complex 1 participates in the assembly of hepatitis C virus through an interaction with NS2 (2011)
  • Author(s): Suzuki R, Suzuki T, Saito K, Matsuda M, Watashi K, Matsuura Y, Wakita T, Aizaki H
  • Organizer: 18th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Seattle, WA, USA
• [Presentation] Identification of host factor that interacts with hepatitis C virus NS2protein and is involved in the viral assembly (2011)
  • Author(s): Suzuki R, Suzuki T, Saito K, Matsuda M, Watashi K, Matsuura Y, Wakita K, Aizaki H
  • Organizer: XV International Congress of Virology
  • Place of Presentation: Sapporo, Japan
• [Presentation] Identification of small molecules affecting late steps of hepatitis C virus life cycle (2011)
  • Author(s): Watashi K, Uchida N, Suzuki R, Aizaki H, Wakita T
  • Organizer: XV International Congress of Virology
  • Place of Presentation: Sapporo, Japan
• [Presentation] Antiviral activity of glycyrrhizin against hepatitis C virus in vitro (2011)
  • Author(s): Matsumoto Y, Watashi K, Suzuki R, Matsuura Y, Suzuki T, Miyamura T, Wake K, Wakita T, Aizaki H
  • Organizer: XV International Congress of Virology
  • Place of Presentation: Sapporo, Japan
• [Presentation] Identification of lipid droplet-associated membrane proteins that are involved in HCV production (2011)
  • Author(s): Aizaki H, Matsumoto Y, Goto K, Watashi K, Suzuki R, Fukasawa M, Hanada K, Sato S, Takahashi N, Matsuura Y, Motojima K, Miyamura T, Suzuki T, Wakita T
  • Organizer: 第18回C型肝炎ウイルス及び関連ウイルスに関する国際研究集会
  • Place of Presentation: アメリカ、シアトル
• [Presentation] Identification and functional analysis of small molecules in hibiting the late step of hepatitis C virus life cycle (2011)
  • Author(s): Watashi K, Uchida N, Suzuki R, Aizaki H, Wakita T
  • Organizer: 第18回C型肝炎ウイルス及び関連ウイルスに関する国際研究集会
  • Place of Presentation: アメリカ、シアトル
• [Presentation] Small molecules that elicit strong anti-HCV activity through down-modulation of HCV entry receptor (2011)
  • Author(s): Takebe Y, Uenishi R, Tani H, Suzuki R, Takagi M, Hase S, Liao H, Tsuchiura T, Shinya K, Wakita T, Matsuura Y, Patel A
  • Organizer: 第18回C型肝炎ウイルス及び関連ウイルスに関する国際研究集会
  • Place of Presentation: アメリカ、シアトル
• [Presentation] Isolation of a highly infectious hepatitis C virus with adaptive mutations (2011)
  • Author(s): Fukasawa M, Shirasago Y, Saito K, Murakami Y, Fukazawa H, Suzuki T, Suzuki R, Wakita T, Hanada K, Chiba J
  • Organizer: 第18回C型肝炎ウイルス及び関連ウイルスに関する国際研究集会
  • Place of Presentation: アメリカ、シアトル
• [Presentation] Identification of novel NS5A-associated proteins in the host-cell membrane fraction and their role in HCV life cycle (2011)
  • Author(s): Goto K, Kimura T, Watashi K, Suzuki R, Yamagoe S, Miyamura T, Moriya K, Yotsuyanagi H, Koike K, Suzuki T, Wakita T, Aizaki H
  • Organizer: 第18回C型肝炎ウイルス及び関連ウイルスに関する国際研究集会
  • Place of Presentation: アメリカ、シアトル
• [Presentation] Halopemide inhibited a post-assembly step in hepatitis C virus life cycle (2011)
  • Author(s): Uchida N, Watashi K, Suzuki R, Aizaki H, Chiba J, Wakita T
  • Organizer: 第18回C型肝炎ウイルス及び関連ウイルスに関する国際研究集会
  • Place of Presentation: アメリカ、シアトル
• [Presentation] Signal peptidase complex 1 participates in the assembly of hepatitis C virus through an interaction with NS2 (2011)
  • Author(s): Suzuki R, Suzuki T, Saito K, Matsuda M, Watashi K, Matsuura Y, Wakita T, Aizaki H
  • Organizer: 第18回C型肝炎ウイルス及び関連ウイルスに関する国際研究集会
  • Place of Presentation: アメリカ、シアトル
• [Presentation] Identification of host factor that interacts with hepatitis C virus NS2 protein and is involved in the viral assembly (2011)
  • Author(s): Suzuki R, Suzuki T, Saito K, Matsuda M, Watashi K, Matsuura Y, Wakita T, Aizaki H
  • Organizer: 第15回国際ウイルス学会
  • Place of Presentation: 札幌
• [Presentation] Identification of small molecules affecting late steps of he patitis C virus life cycle (2011)
  • Author(s): Watashi K, Uchida N, Suzuki R, Aizaki H, Wakita T
  • Organizer: 第15回国際ウイルス学会
  • Place of Presentation: 札幌
• [Presentation] Antiviral activity of glycyrrhizin against hepatitis C virus in vitro (2011)
  • Author(s): Matsumoto Y, Watashi K, Suzuki R, Matsuura Y, Suzuki T, Miyamura T, Wake K, Wakita T, Aizaki H
  • Organizer: 第15回国際ウイルス学会
  • Place of Presentation: 札幌
• [Presentation] Entry mechanisms of hepatitis C virus examined by trans-complemented particles (2010)
  • Author(s): Suzuki R, Alazawa D, Ishii K, Matsuura Y, Wakita T, Suzuki T
  • Organizer: 9th International Symposium on Positive-Strand RNA Viruses
  • Place of Presentation: Atlanta, GA, USA
• [Presentation] Efficient production of trans-complemented hepatitis C virus particles : Use for study of viral entry process (2010)
  • Author(s): Suzuki R, Alazawa D, Ishii K, Matsuura Y, Wakita T, Suzuki T
  • Organizer: 17th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Yokohama, Japan
• [Presentation] C型肝炎ウイルスのtrans-packaging型粒子を用いた感染機構の解析 (2010)
  • Author(s): 鈴木亮介、斎藤憲司、赤澤大輔、石井孝司、松浦善治、脇田隆字、鈴木哲朗
  • Organizer: 日本ウイルス学会第58回学術集会
  • Place of Presentation: 徳島
• [Presentation] 分割ユビキチン法を利用したHCV NS2と結合する宿主因子の探索およびウイルス粒子形成への関与 (2010)
  • Author(s): 鈴木亮介、相崎英樹、脇田隆字、鈴木哲朗
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 神戸市
• [Presentation] Entry mechanisms of hepatitis C virus examined by trans-complemented particles (2010)
  • Author(s): Ryosuke Suzuki, Daisuke Akazawa, Koji Ishii, Yoshiharu Matsuura, Takaji Wakita, Tetsuro Suzuki
  • Organizer: 第9回プラス鎖RNAウイルスに関する国際研究集会
  • Place of Presentation: アメリカ、アトランタ
• [Presentation] Efficient production of traps-complemented hepatitis C virus particles : Use for study of viral entry process (2010)
  • Author(s): Ryosuke Suzuki, Daisuke Akazawa, Koji Ishii, Yoshiharu Matsuura, Takaji Wakita, Tetsuro Suzuki
  • Organizer: 第17回C型肝炎ウイルス及び関連ウイルスに関する国際研究集会
  • Place of Presentation: 横浜市
• [Presentation] C型肝炎ウイルスのtrans-packaging型粒子を用いた感染機構の解析 (2010)
  • Author(s): 鈴木亮介、斎藤憲司、赤澤大輔、石井孝司、松浦善治、脇田隆字、鈴木哲朗
  • Organizer: 第58回日本ウイルス学会学術集会
  • Place of Presentation: 徳島市
• [Presentation] Involvement of PA28gamma and E6AP in the degradation of HCV core protein and the viral production (2009)
  • Author(s): Moriishi K, Shoji I, Suzuki R, Suzuki T, Matsuura Y
  • Organizer: 16th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Nice, France
• [Presentation] Plasmid-based production of trans-complemented HCV particles : its use for functional analysis of NS2 (2009)
  • Author(s): Suzuki R, Saito K, Ando T, Ishii K, Matsuura Y, Miyamura T, Wakita T, Suzuki T
  • Organizer: 16th International Meeting on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Nice, France
• [Presentation] HCVコア蛋白質のプロテアソームによる分解とウイルス産生制御 (2009)
  • Author(s): 森石恒司、勝二郁夫、鈴木亮介、鈴木哲朗、松浦善治
  • Organizer: 日本ウイルス学会第57回学術集会
  • Place of Presentation: 東京
• [Presentation] C型肝炎ウイルスのtrans-packaging系を用いたNS2蛋白質の感染性粒子形成における機能解析 (2009)
  • Author(s): 鈴木亮介、斎藤憲司、安東友美、石井孝司、松浦善治、宮村達男、脇田隆字、鈴木哲朗
  • Organizer: 日本ウイルス学会第57回学術集会
  • Place of Presentation: 東京
• [Presentation] Prohibitin2はC型肝炎ウイルス(HCV) NS5A蛋白と相互作用しHCV複製調節に働く (2009)
  • Author(s): 木村敬郎、鈴木亮介、山越智、鈴木健裕、堂前直、勝二郁夫、松浦善治、千葉丈、脇田隆字、鈴木哲朗
  • Organizer: 日本ウイルス学会第57回学術集会
• [Presentation] Plasmid-based production of trans-complemented HCV particles : its use for functional analysis of NS2 (2009)
  • Author(s): 鈴木亮介, 斎藤憲司, 安東友美, 石井孝司, 松浦善治, 宮村達男, 脇田隆字, 鈴木哲朗
  • Organizer: 第16回C型肝炎ウイルス及び関連ウイルスに関する国際研究集会
  • Place of Presentation: フランス、ニース
• [Presentation] C型肝炎ウイルスNS2蛋白質が感染性粒子形成に関与するメカニズムの解明 (2009)
  • Author(s): 鈴木亮介、斎藤憲司、安東友美、石井孝司、松浦善治、宮村達男、脇田隆字、鈴木哲朗
  • Organizer: 第57回日本ウイルス学会学術集会
  • Place of Presentation: 東京
• [Patent(Industrial Property Rights)] C型肝炎ウイルス粒子高生産系
  • Inventor(s): 脇田隆字、石井孝司、鈴木哲朗、鈴木亮介、宮村達男、田邉純一、曽根三郎
  • Overseas
• [Patent(Industrial Property Rights)] 新規組換え型ヒトC型肝炎ウイルス様粒子とその産生方法
  • Inventor(s): 脇田隆字、石井孝司、鈴木哲朗、鈴木亮介、宮村達男、田邉純一、曽根三郎
  • Overseas