The study of molecular mechanism of hepatitis C virusproduction

2012 Fiscal Year Final Research Report

• Project/Area Number: 22590731
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Kobe University
• Principal Investigator: SHOJI Ikuo 神戸大学, 大学院・医学研究科, 准教授 (40356241)
• Co-Investigator(Kenkyū-buntansha): IDE Yoshihiro 神戸大学, 大学院・医学研究科, 助教 (70258606)
• Project Period (FY): 2010 – 2012
• Keywords: 肝臓学

Research Abstract

Development of HCV replicon cells and HCV production system using HCV JFH1 strain enabled us to investigate the whole life cycle of HCV and the host factors essential for HCV production. In this study, we aimed to clarify molecular mechanism of HCV production by investigating binding partners for HCV proteins. We analyzed HCV core-binding proteins, hnRNPH1/H2/F. Addition of HCV RNA inhibited the binding between the core protein and hnRNPs. RNase recovered this inhibition. Theseresults suggest that RNA is required for the interaction between the core protein and hnRNPs. Our results suggest that these hnRNPH and hnRNPF cooperatively regulate HCV replication. We then identified HCV NS5A-binding proteins, ubiquitin ligase p138, transcription factor HNF-1a, and histone methyltransferase SMYD3. The p138 protein bound NS5A protein, but not other HCV proteins. Our data suggest that p138 protein promotes ubiquitylation of NS5A protein and enhances its proteasomal degradation, suggesting that p138 regulates HCV replication. We analyzed the p138-binding domain on the NS5A protein using a series of deletion mutants. Then weanalyzed the ubiquitylation site on the NS5A protein using 13 point-mutants. These studies would contribute to the better understanding of HCV life cycles as well as the viral pathogenesis.

Research Products

• [Journal Article] Roles of the two distinct proteasome pathways in hepatitis C virus infection (2012)
  • Author(s): Shoji, I.
  • Journal Title: World Journal of Virology Volume: 1 Pages: 44-50
  • Peer Reviewed
• [Journal Article] Hepatitits C virus infection suppresses GLUT2 gene expression via down-regulation of hepatocyte nuclear factor 1α (2012)
  • Author(s): Matsui, C., Shoji, I., Kaneda, S., Sianipar, IR., Deng, L., and Hotta, H
  • Journal Title: Journal of Virology Volume: 86(23) Pages: 12903-11
  • Peer Reviewed
• [Journal Article] K. HCV NS5A protein containing potential ligands for both Src homology 2 and 3 domains enhances autophosphorylation of Src family kinases Fyn in B cells (2012)
  • Author(s): Nakashima, K., Takeuchi, K., Chihara, K., Deng, L., Shoji, I., Hotta, H., and Sada
  • Journal Title: PLoS One Volume: 7(10) Pages: e46634
  • Peer Reviewed
• [Journal Article] Molecular mechanism of hepatitis C virus-induced glucose metabolic disorders (2012)
  • Author(s): Shoji, I., Deng L., and Hotta, H
  • Journal Title: Frontiers in Microbiology Volume: 2 Pages: 1-5
  • Peer Reviewed
• [Journal Article] Generation of a recombinant reporter hepatitis C virus useful for the analyses of virus entry, intra-cellular replication and virion production (2012)
  • Author(s): Kamada, K., Shoji, I., Deng, L, Wakita, T., and Hotta, H
  • Journal Title: Microbes and Infection Volume: 14 Pages: 69-78
  • Peer Reviewed
• [Journal Article] A point mutation at Asn-534 that disrupts a conserved N-glycosylation motif of the E2 glycoprotein of hepatitis C virus markedly enhances the sensitivity to antibody neutralization. (2012)
  • Author(s): Sasayama, M., Shoji, I., Adianti M, Jiang DP, Deng, L, Saito T, Watanabe H, Kawata S, Aoki C, and Hotta, H
  • Journal Title: Journal of Medical Virology Volume: 84 Pages: 229-34
  • Peer Reviewed
• [Journal Article] Polymorphisms of hepatitis C virus NS5A and core proteins and clinical outcome of pegylated interferon/ribavirin therapy (2012)
  • Author(s): El-Shamy, A., Ide, Y-H., Kim, SR., Sasase, N., Imoto, S., Deng, L., Shoji, I., and Hotta, H.
  • Journal Title: Intervirology Volume: 55 Pages: 1-11
  • Peer Reviewed
• [Journal Article] Sequence heterogeneity in NS5A of hepatitis C virus gentypes 2a and 2b and clinical outcome of pegylated-Interferon/Ribavirin therapy (2012)
  • Author(s): El-Shamy, A., Shoji, I., Kim S-R., Ide, Y-H., Imoto, S., Deng, L., Yoon, S., Fujisawa, T., Tani, S., Yano, Y., Seo, Y., Azuma, T., and Hotta, H
  • Journal Title: PLoS One Volume: 7 Pages: 1-10
  • Peer Reviewed
• [Journal Article] Sequence heterogeneity of NS5A and core proteins of hepatitis C virus and virological responses to Pegylated-Interferon/Ribavirin combination therapy (2011)
  • Author(s): El-Shamy, A., Shoji, I., Saito, T., Watanabe H., Ide, Y-H., Deng, L., Kawata, S., and Hotta, H.
  • Journal Title: Microbiology and Immunology Volume: 55 Pages: 418-26
  • Peer Reviewed
• [Journal Article] Hepatitis C virus infection promotes hepatic gluconeogenesis through an NS5A-mediated, FoxO1-dependent pathway (2011)
  • Author(s): Deng, L., Shoji, I., Oawa, W., Kaneda, S., Soga, T., Jiang, D. P., Ide, Y-H., and Hotta, H
  • Journal Title: Journal of Virology Volume: 85 Pages: 8556-68
  • Peer Reviewed
• [Journal Article] Chaperonin TRiC/CCT participates in replication of hepatitis C virus genome via interaction with the viral NS5B protein (2011)
  • Author(s): Inoue, Y., Aizaki, H., Hara, H., Matsuda, M., Ando, T., Shimoji, T., Murakami, K., Masaki, T., Shoji, I., Homma, S., Matsuura, Y., Miyamura, T., Lai, MMC, Wakita, T., and Suzuki, T
  • Journal Title: Virology Volume: 410 Pages: 38-47
  • Peer Reviewed
• [Journal Article] Estradiol inhibits the production of infectious particles of hepatitis C virus (2010)
  • Author(s): Hayashida K, Shoji, I., Deng, L., Ide, Y-H., and Hotta, H. 17
  • Journal Title: Microbiology and Immunology Volume: 54 Pages: 684-90
  • Peer Reviewed
• [Journal Article] E6AP ubiquitin ligase mediates ubiquitin-dependent degradation of peroxiredoxin 1 (2010)
  • Author(s): Nasu, J., Murakami, K., Miyagawa, S., Yamashita, R., Ichimura, T., Wakita, T., Hotta, H., Miyamura, T., Suzuki, T., Satoh, T., and Shoji, I.
  • Journal Title: Journal of Cellular Biochemistry Volume: 111 Pages: 676-85
  • Peer Reviewed
• [Journal Article] Involvement of PA28γ 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 Pages: 411-420
  • Peer Reviewed
• [Journal Article] Secondary structure of the amino-terminal region of HCV NS3 and virological response to pegylated interferon plus ribavirin therapy for chronic hepatitis C (2010)
  • Author(s): Sanjo M., Saito, T., Ishii, R., Nishise, Y., Haga, H., Okumoto, K., Ito, J., Watanabe, H., Saito, K., Togashi, H., Fukuda, K., Imai, Y., El-Shamy, A., Deng, L., Shoji, I., Hak, H., and Kawata, S
  • Journal Title: Journal of Medical Virology Volume: 82 Pages: 1364-70
  • Peer Reviewed
• [Presentation] Molecular mechanism of hepatitis C virus-induced glucose metabolism disorder. (2012)
  • Author(s): Shoji I
  • Organizer: The 10th JSH single topic conference
  • Place of Presentation: 東京
  • Year and Date: 20121121-22
• [Presentation] C 型肝炎ウイルス NS5A 蛋白質の新規結合因子ヒストンメチル基転移酵素SMYD3 の同定と機能解析 (2012)
  • Author(s): 陳明、甘翔、Deng Lin、勝二郁夫、堀田博
  • Organizer: 第60回日本ウイルス学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 20121113-15
• [Presentation] C 型肝炎ウイルスによるGLUT2 遺伝子発現抑制の分子機構 (2012)
  • Author(s): 松井千絵子、勝二郁夫、Deng Lin、堀田 博
  • Organizer: 第60回日本ウイルス学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 20121113-15
• [Presentation] HCV infection induces lysosomal degradation of hepatocyte nuclear factor 1α via interaction with HCV NS5A protein (2012)
  • Author(s): Matsui C, Shoji I, Deng L, Hotta H
  • Organizer: 19th International Symposium on Hepatitis C Virus and Related Viruses
  • Place of Presentation: Venice, Italy
  • Year and Date: 20121005-09
• [Presentation] Identification and characterization of a novel NS5A-interacting protein (2012)
  • Author(s): Chen M, Gan X, Deng L, Shoji I, Hotta H
  • Organizer: 19th International Symposium on Hepatitis C Virus and Related Viruses.
  • Place of Presentation: Venice, Italy
  • Year and Date: 20121005-09
• [Presentation] Identification of an E3 ubiquitin ligase that mediates ubiquitylation of hepatitis C virus NS5A protein (2011)
  • Author(s): Shoji I, Okada N, Gan X, Miyagawa S, Makimoto M, Chieko M, Jang DP, Deng L, Ide Y-H, andHotta H
  • Organizer: 第34回日本分子生物学会年
  • Place of Presentation: 横浜
  • Year and Date: 2011-12-14
• [Presentation] C 型肝炎ウイルス NS5A の新規結合タンパク質であるヒストンメチル基転移酵素SMYD3 の同定 (2011)
  • Author(s): 甘翔、Lin Deng、陳明、井出良浩、勝二郁夫、堀田博
  • Organizer: 第64回日本細菌学会関西支部総会
  • Place of Presentation: 大阪
  • Year and Date: 2011-11-19
• [Presentation] Identification of an E3 ubiquitin ligase that mediates ubiquitylation of hepatitis C virus NS5A protein (2011)
  • Author(s): Shoji I, Okada N, Gan X, Miyagawa S, Makimoto M, El-Shamy A, Deng L, Jang DP, Ide Y-H, and Hotta H
  • Organizer: International Union of Microbiological Societies 2011 Congress
  • Place of Presentation: 札幌
  • Year and Date: 2011-09-16
• [Presentation] Identification of an E3 ubiquitin ligase that targets hepatitis C virus NS5A protein for ubiquitylation. (2011)
  • Author(s): Shoji I, Okada N, Gan X, Miyagawa S, MakimotoM, El-Shamy A, Deng L, Jang DP, IdeY-H, and Hotta H
  • Organizer: 18thInternational symposium on hepatitis C virus and related viruses
  • Place of Presentation: Seattle, USA
  • Year and Date: 2011-09-10
• [Presentation] C型肝炎ウイルスのインターフェロン・リバビリン治療応答性を規定するウイルス側因子 (2011)
  • Author(s): 勝二郁夫、Ahmed El-Shamy, 堀田博
  • Organizer: 第2回神戸肝炎ウイルス学術講演会
  • Place of Presentation: 神戸
  • Year and Date: 2011-07-22
• [Presentation] E6AP uibiquitin ligase mediatesubiquitin-dependent degradation of peroxiredoxin 1 (2010)
  • Author(s): Shoji I, Nasu J, Murakami K, Miyagawa S, Yamashita R, IchimuraT, Wakita T, Ide Y-H, Hotta H, Miyamura T, Suzuki T,Satoh T
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 20101207-10
• [Presentation] C型肝炎ウイルスNS5Aに結合するユビキチンリガーゼの同定 (2010)
  • Author(s): 岡田典子、勝二郁夫、甘翔、Deng L、姜大鵬、井出良浩、堀田博
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 20101207-10
• [Presentation] HCVによる糖代謝障害の分子機序 (2010)
  • Author(s): 勝二郁夫、Deng L、堀田博
  • Organizer: 第58回日本ウイルス学会学術集会
  • Place of Presentation: 徳島
  • Year and Date: 20101107-09
• [Presentation] 糖代謝に及ぼすC型肝炎ウイルスの影響及びその分子機序の解析 (2010)
  • Author(s): Deng L, 兼田崇作, 井出良浩, 勝二郁夫,堀田博
  • Organizer: 第58回日本ウイルス学会学術集会
  • Place of Presentation: 徳島
  • Year and Date: 20101107-09
• [Presentation] Molecular mechanism of HCVinduced suppression of glucose transporter(GLUT)2 expression. (2010)
  • Author(s): Shoji I, Kaneda S, Deng L, Ide Y-H,Hotta H.
  • Organizer: 17thInternational Meeting on hepatitis C virus and related viruses
  • Place of Presentation: Yokohama, Kanagawa
  • Year and Date: 20100910-14
• [Presentation] HCV-induced generation of reactive oxygen species leads to Bax-mediated apoptosis through activation of the c-Jun NH_2-terminal kinase signaling pathway. (2010)
  • Author(s): Deng L, Ide YH, Shoji I, Hotta H
  • Organizer: 17th International Meeting on hepatitis C virus and related viruses.
  • Place of Presentation: Yokohama, Kanagawa.
  • Year and Date: 20100910-14
• [Presentation] C型肝炎ウイルス感染は糖新生系を亢進し、糖尿病発症に関与する (2010)
  • Author(s): 堀田博、井出良浩、勝二郁夫
  • Organizer: 第46回日本肝臓学会総会
  • Place of Presentation: 山形
  • Year and Date: 20100527-28
• [Presentation] Role of the proteasome pathways in the hepatitis C virus life cycle (2010)
  • Author(s): Shoji I
  • Organizer: RCC-ERI Seminar
  • Place of Presentation: National Institute of Health, Bankok, Thailand.
  • Year and Date: 2010-08-08
• [Book] 最新!C 型肝炎治療薬の使いかた. (2012)
  • Author(s): 勝二郁夫
  • Total Pages: 83-87
  • Publisher: 診断と治療社, 岡上武監修
• [Book] シンプル微生物学 (2011)
  • Author(s): 勝二郁夫
  • Total Pages: 247-252
  • Publisher: 南江堂
• [Book] はじめの一歩のイラスト感染症・微生物学 (2011)
  • Author(s): 勝二郁夫
  • Total Pages: 79-82
  • Publisher: 羊土社
• [Remarks]
  • URL: http://www.med.kobe-u.ac.jp/micro/1.htm