The analysis of the DNA damage and genetic repair mechanism in Hepatitis C

• Project/Area Number: 19590419
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Experimental pathology
• Research Institution: Tokyo Metropolitan Organization for Medical Research
• Principal Investigator: NURIYA Hideko Tokyo Metropolitan Organization for Medical Research, 東京都臨床医学総合研究所, 主任研究員 (80270685)
• Co-Investigator(Kenkyū-buntansha): KOHARA Michinori 財団法人東京都医学研究機構, 東京都臨床医学総合研究所, 参事研究員 (10250218)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2008: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2007: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 細胞 / HCV / DNA傷害 / 修復異常 / 修復関連蛋白 / 肝細胞培養系 / 局在変化

Research Abstract

HCVによる遺伝子傷害または遺伝子修復機能に与える影響を解析するため、DNA二本鎖切断(DSB)を導入後に相同組換え(修復)が起こるとGFPの蛍光を発生する特徴を持つヒト繊維肉腫細胞株に、レンチウイルスでHCV各蛋白を発現させ35日間培養した。この各HCV蛋白発現細胞を用いて、経日的なFACS解析によりGFPの蛍光を生じた細胞数の割合から相同組換え(修復)の異常の有無を確認した。強制的にDNA傷害を誘導する系と誘導しない系の両者で実施した。
各実験系でGFP陽性細胞の割合がEmpと差があるHCV蛋白が存在し、HCV蛋白によるDNA修復異常の可能性が示唆された。

Research Products

• [Journal Article] Modification of hepatitis C virus 1b RNA polymerase to make a highly active JFH1-type polymerase by mutation of the thumb domain (2009)
  • Author(s): Leiyun Weng Jiamu Du Jingling Zhou Jianping Ding Takaji Wakita Michinori Kohara Tetsuya Toyoda.
  • Journal Title: Arch Virol. in press Pages: 0-0
  • Peer Reviewed
• [Journal Article] Disruption of IFN Signaling and HCV Synergistically Enhance Lymphoproliferation through Type II CD95 and Interleukins. (2009)
  • Author(s): Keigo Machida, Kyoko Tsukiyama-Kohara, Eiji Seike, Shigenobu Tone, Yukiko Hayashi, Yuri Kasama, Masumi Shimizu, Hidemi Takahashi, Chyoji Taya, Hiromichi Yonekawa, Nobuyuki Tanaka, and Michinori Kohara.
  • Journal Title: Gastroenterology in press Pages: 0-0
  • Peer Reviewed
• [Journal Article] Synthesis of nuclease- resistant siRNAs possessing universal overhangs. (2009)
  • Author(s): Yoshihito Ueno, Yuuji Watanabe, Aya Shibata, Kayo Yoshikawa, Takashi Takano,Michinori Kohara, Yukio Kitade.
  • Journal Title: Bioorganic & Medicinal Chemistry in press Pages: 0-0
  • Peer Reviewed
• [Journal Article] A mutational shift from domain III to II in the internal ribosome entry site of hepatitis C virus after interferon-ribavirin therapy. (2008)
  • Author(s): Kei Ogata, Takahito Kashiwagi, Jun Iwahashi, Koyu Hara, Haruhito Honda, Tatsuya Ide, Ryukichi Kumashiro, Michinori Kohara, Michio Sata, and Nobuyuki Hamada.
  • Journal Title: Arch Virol. 153(8) Pages: 1575-9
  • Peer Reviewed
• [Journal Article] Inhibition of hepatitis C virus infection and expression in vitro and in vivo by recombinant adenovirus expressing short hairpin RNA. (2008)
  • Author(s): Naoya Sakamoto, Yoko Tanabe, Takanori Yokota, Kenichi Satoh, Yuko Sekine-Osajima, Mina akagawa, Yasuhiro Itsui, Megumi Tasaka, Yuki Sakurai, Chen Cheng-Hsin, Masahiko Yano, Shogo Ohkoshi, Yutaka Aoyagi, Shinya Maekawa, Nobuyuki Enomoto, Michinori Kohara, and Mamoru Watanabe.
  • Journal Title: J. Gastroenterology Hepatology. 23(9) Pages: 1437-47
  • Peer Reviewed
• [Journal Article] Comparative aspects on the role of polypyrimidine tract-binding protein in internal initiation of hepatitis C virus and picornavirus RNAs. (2008)
  • Author(s): T. Nishimura, M. Saito, T. Takano, A. Nomoto, M. Kohara, K. Tsukiyama-Kohara.
  • Journal Title: Compara. Immu. Microbio. Infect. Dises. 31 Pages: 435-448
  • Peer Reviewed
• [Journal Article] Hepatitis C virus NS5A protein interacts with and negatively regulates the non-receptor protein-tyrosine kinase Syk. (2008)
  • Author(s): Sachiko Inubushi, Motoko Nagano-Fujii, Kikumi Kitayama, Motofumi Tanaka, Chunying An, Hiroshi Yokozaki, Hirohei Yamamura, Hideko Nuriya, Michinori Kohara, Kiyano Sada, and Hak Hotta.
  • Journal Title: J. Gen. Virology 89 Pages: 1231-1242
  • Peer Reviewed
• [Journal Article] Hepatitis C virus NS5A protein interacts with and negatively regulates the non-receptor protein-tyrosine kinase Syk (2008)
  • Author(s): Sachiko Inubushi, Motoko Nagano-Fujii, Kikumi Kitayama
  • Journal Title: Journal of General Virology 89 Pages: 1231-1242
  • Peer Reviewed
• [Journal Article] Therapeutic application of RNA interference for Hepatitis C Virus. (2007)
  • Author(s): Tsunamasa Watanabe, Takuya Umehara, Michinori Kohara.
  • Journal Title: Advanced Drug Delivery Reviews 59 Pages: 1263-1276
• [Journal Article] Liver target delivery of small interfering RNA to the HCV gene by lactosylated cationic liposome. (2007)
  • Author(s): Tsunamasa Watanabe, Takuya Umehara, Fumihiko Yasui, Shin-ichiro Nakagawa, Junichi Yano, Tadaaki Ohgi, Satoru Sonoke, Kazuaki Inoue, Makoto Yoshiba and Michinori Kohara.
  • Journal Title: J. Hepatology 47 Pages: 744-750
• [Journal Article] Survival of liver failure pigs by transplantation of reversibly immortalized human hepatocytes with Tamoxifen-mediated self-recombination. (2007)
  • Author(s): Totsugawa T, Yong C, Rivas-Carrillo JD, Soto-Gutierrez A, Navarro-Alvarez N, Noguchi H, Okitsu T, Westerman KA, Kohara M, Reth M, Tanaka N, Leboulch P, Kobayashi N.
  • Journal Title: J. Hepatology 47 Pages: 74-82
  • Peer Reviewed
• [Journal Article] Identification of CTL epitopes in hepatitis C virus by a genome-wide computational scanning and a rational design of peptide vaccine. (2007)
  • Author(s): Toshie Mashiba, Keiko Udaka, Yasuko Hirachi, Yoichi Hiasa, Tomoya Miyakawa, Yoko Satta, Tsutomu Osoda, Sayo Kataoka, Michinori Kohara and Morikazu Onji.
  • Journal Title: Immunogenetics 59 Pages: 197-209
  • Peer Reviewed
• [Journal Article] Evaluation of a Cyclophilin Inhibitor in Hepatitis C Virus-Infected Chimeric Mice In Vivo. (2007)
  • Author(s): Kazuaki Inoue, Takuya Umehara, Urs T. Ruegg, Fumihiko Yasui, Tsunamasa Watanabe, Hiroshi Yasuda, Makoto Yoshiba, Jean-Maurice Dumont, Pietro Scalfaro, Michinori Kohara.
  • Journal Title: Hepatology 45 Pages: 921-928
  • Peer Reviewed
• [Journal Article] Inhibition of Hsp90 suppresses HCV replication in replicon cell lines and in chimeric mice with humanized liver. (2007)
  • Author(s): Shin-ichiro Nakagawa, Takuya Umehara, Chiho Matsuda, Shusuke Kuge, Masayuki Sudoh and Michinori Kohara.
  • Journal Title: Biochem. Biophys. Res. Commun. 353 Pages: 882-888
  • Peer Reviewed
• [Presentation] Serine palmitoyltransferase inhibitor suppresses HCV replication in a mouse model. (2008)
  • Author(s): Hirata Y., Umehara T., Sudoh M., Kohara M.
  • Organizer: XIV International Congress of Virology
  • Place of Presentation: Istanbul
• [Presentation] Serine Palmitoyltransferase Inhibitor Suppresses HCV Replication in a Mouse Model. (2008)
  • Author(s): Hirata Y. Umehara T., Sudoh M., Yasui F., Kohara M.
  • Organizer: 21th International Conference on Antiviral Research
  • Place of Presentation: Montreal
• [Presentation] Characterization of HCV Replication regulatory by DHCR24. (2007)
  • Author(s): Takano T., Hayashi M., Umehara T., Hirata Y., Tsukiyama-Kohara K., Kohara M.
  • Organizer: 第30回日本分子生物学会
  • Place of Presentation: 横浜
• [Presentation] The novel regulatory pathway of TOM70 concerning with cell death. (2007)
  • Author(s): Takano T., Kohara M., Kai C., Tsukiyama-Kohara K.
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
• [Remarks]
  • URL: http://www.rinshoken.or.jp/
• [Remarks]
  • URL: http://www.rinshoken.or.jp/