2000 Fiscal Year Annual Research Report

C型肝炎ウイルス(HCV)NS5BのRNA合成活性と炎症応答の修飾

Research Project

Project/Area Number	11694257
Research Institution	KANAZAWA UNIVERSITY
Principal Investigator	村上清史金沢大学, がん研究所, 教授 (90019878)
Co-Investigator(Kenkyū-buntansha)	林直之金沢大学, がん研究所, 助手 (50253456) 善岡克次金沢大学, がん研究所, 助教授 (60200937) 須田貴司金沢大学, がん研究所, 教授 (70250090) 野村孝弘金沢大学, がん研究所, 助手 (80115261)
Keywords	HCV NS5B / RNA依存RNA合成酵素(RdRP) / アラニンスキャニング / RNA結合 / HCV惑染 / NF-kB活性化
Research Abstract	HCVの持続感染の遮断が肝がん発症のリスクを低下させると期待される。HCVは宿主とは異なるRNA複製過程があり、RNA複製酵素であるNS5BはHCV複製遮断の有力な標的である。HCV複製の阻止を目標に、HCV複製酵素NS5Bの構造と機能の検討と、NS5Bの宿主の炎症応答経路への影響の検討を行った。我々は、集約的な及び1残基のアラニン置換変異を系統的に導入し、NS5Bの諸機能を解析した。その結果1.NS5BのRNA依存RNA合成(RdRP)活性に必須である5アミノ酸残基を新たに特定した(Hepatology,in press)。2.NS5BのRdRP活性には、鋳型結合能は要求せず、鋳型/primer結合能を要求した。Y276が鋳型/primerに必須なアミノ酸残基として特定された。3.活性中心より離れたRdRP活性に必須な2残基が、NS5Bのhomomericな相互作用に関わることを見いだし、各種の置換残基を用いてhomomericな作用がRdRP活性に必須で、この2残基は相互作用に直接関わる残基の可能性が強く示唆された。4.NS5Bの過剰発現系で炎症関連シス配列AP-1,NFkBの活性化を認めた。しかしアラニンスキャンでNS5Bの特定配列との相関が認められず、発現量によってNS5Bの転写活性化効果は変動した。NS5Bによる炎症関連遺伝子の修飾の可能性はあるが、感染時にHCVの他の蛋白が同時発現する条件で、NS5Bの微量発現が今回観察された効果が得られるか否かは今後の検討に残された。

Research Products
(23 results)

All Other

All Publications (23 results)

[Publications] Tul,DY., et al.: "Incidence of hepatocellular carcinoma in the transgenic mice expressing hepatitis B virus X-protein."J.Hepatol.. 31. 123-132 (1999)
[Publications] Nomura,T., et al.: "Human Hepatitis B Virus X protein is detectable in nuclei of transfected cells, And active for transactivation."Biochim.Biophys.Acta.. 1453. 330-340 (1999)
[Publications] Ohno,H., et al.: "Human Hepatitis B Virus X protein augments the DNA binding of nuclear Factor for IL & through its basic leucine zipper domain."J.Med.Viol.. 58. 11-18 (1999)
[Publications] Murakami,S.: "Hepatitis B Virus X protein : Structure, Functions and Biological Roles."Intervirology. 42. 81-99 (1999)
[Publications] 村上清史: "B型肝炎ウイルスX蛋白の発現と機能"生化学. 71.11. 1309-1326 (1999)
[Publications] Kuwano,K., et al.: "Essential roles of the Fas-Fas ligand pathway in the development of pulmonary fibrosis."J.Clin.Invest.. 104. 13-19 (1999)
[Publications] Koyano,S., et al.: "A novel Jun N-terminal kinase (JNK)-binding protein that enhances the activation of JNK by MEK kinase 1 and TGF-activated kinase 1"FEBS Lett.. 457. 385-388 (1999)
[Publications] Ito,M.Et al.: "JSAP1, a novel Jun N-terminal kinase (JNK) that functions as a scaffold factor in the JNK signaling pathway."Mol.Cell.Biol.. 19. 7539-7548 (1999)
[Publications] Takao,N., et al.: "Disruption of ATM in p53-null cells causes multiple functional abnormalities in celllar response to ionizing radiation."Oncogene. 18. 7002-7009 (1999)
[Publications] Zhang,Y., et al.: "Immunosuppressant FK506 activates NFkB through the proteasome-mediated degradation of IkB"J.Biol.Chem.. 274. 34657-34662 (1999)
[Publications] Miwa,K., et al.: "Therapeutic of an anti-Fas ligand mAb on lethal graftversus-host disease."Int.Immunol.. 11. 925-931 (1999)
[Publications] Saito,I., et al.: "Fas ligand-mrdiated exocrinopathy resembling Sjogren's syndrome in mice transgenic for IL-10."J.Immunol.. 162. 2488-2494 (1999)
[Publications] Ikeda,A., et al.: "P300/CBP-dependent and-independent transcriptional interference between NK-kB RelA and p53."Biochem.Biophys.Res.Communs.. 272. 375-379 (2000)
[Publications] Ito,M., et al.: "Isoforms of JSAP1 scaffold protein generated through alternative splicing."Gene. 255. 229-234 (2000)
[Publications] S-W.Kee, et al.: "Human Hepatitis B Virus X protein is a possible mediator of hypoxia induced angiogenesis in hepatocellular carcinoma."Biochem.Res.Communs.. 268. 456-461 (2000)
[Publications] Dorjsuren,D.: "B型肝炎ウイルス(HBV)Xタンパクに拮抗する新規タンパクRMP。"実験医学. 18. 1137-1141 (2000)
[Publications] K.Masutomi, et. al,.: "Teromerase Activity Reconstituted in Vitro with Purified Human Teromerase Reverse Transcriptase and Human Teromerase RNA Component."J.Biol.Chem.. 275. 22568-22573 (2000)
[Publications] 増富健吉: "昆虫細胞発現系を用いたヒトテロメアーゼ逆転写酵素(hTERT)の発現・精製。"バイオインダストリー. 17. 35-42 (2000)
[Publications] Wei,W., et al.: "Direct interaction between the subunit RAP30 of TFHF and RNA polymeras subunit5 (RPB5) which contributes to the association between TFHF and RNA polymerase II."J.Biol.Chem. (in press). (2001)
[Publications] Kuboki,Y., et al.: "A scaffold protein in the c-Jun NH2-terminal kinase signaling pathways suppresses the extracelller signal-regulated kinase signaling pathway."J.Biol.Chem.. (in press). (2001)
[Publications] Qin,W., et al.: "Mutational Analysisof the Structure and Functions of Hepatitis C Virus RNA-dependent RNA polymerase."Hepatol.. (in press). (2001)
[Publications] 村上清史: "ヒトテロメアーゼ逆転写酵素(hTERT)の発現・精製とヒトテロメアーゼ活性の試験管内の再構成。"教育通信. (印刷中). (2001)
[Publications] 村上清史: "ウイルス肝炎-最新情報「B型肝炎ウイルス(HVB)の遺伝子制御」"富士レビオ株式会社. 271 (1999)

2000 Fiscal Year Annual Research Report

C型肝炎ウイルス(HCV)NS5BのRNA合成活性と炎症応答の修飾

Principal Investigator

村上 清史 金沢大学, がん研究所, 教授 (90019878)

Research Products

[Publications] Tul,DY., et al.: "Incidence of hepatocellular carcinoma in the transgenic mice expressing hepatitis B virus X-protein."J.Hepatol.. 31. 123-132 (1999)

[Publications] Nomura,T., et al.: "Human Hepatitis B Virus X protein is detectable in nuclei of transfected cells, And active for transactivation."Biochim.Biophys.Acta.. 1453. 330-340 (1999)

[Publications] Ohno,H., et al.: "Human Hepatitis B Virus X protein augments the DNA binding of nuclear Factor for IL & through its basic leucine zipper domain."J.Med.Viol.. 58. 11-18 (1999)

[Publications] Murakami,S.: "Hepatitis B Virus X protein : Structure, Functions and Biological Roles."Intervirology. 42. 81-99 (1999)

[Publications] 村上清史: "B型肝炎ウイルスX蛋白の発現と機能"生化学. 71.11. 1309-1326 (1999)

[Publications] Kuwano,K., et al.: "Essential roles of the Fas-Fas ligand pathway in the development of pulmonary fibrosis."J.Clin.Invest.. 104. 13-19 (1999)

[Publications] Koyano,S., et al.: "A novel Jun N-terminal kinase (JNK)-binding protein that enhances the activation of JNK by MEK kinase 1 and TGF-activated kinase 1"FEBS Lett.. 457. 385-388 (1999)

[Publications] Ito,M.Et al.: "JSAP1, a novel Jun N-terminal kinase (JNK) that functions as a scaffold factor in the JNK signaling pathway."Mol.Cell.Biol.. 19. 7539-7548 (1999)

[Publications] Takao,N., et al.: "Disruption of ATM in p53-null cells causes multiple functional abnormalities in celllar response to ionizing radiation."Oncogene. 18. 7002-7009 (1999)

[Publications] Zhang,Y., et al.: "Immunosuppressant FK506 activates NFkB through the proteasome-mediated degradation of IkB"J.Biol.Chem.. 274. 34657-34662 (1999)

[Publications] Miwa,K., et al.: "Therapeutic of an anti-Fas ligand mAb on lethal graftversus-host disease."Int.Immunol.. 11. 925-931 (1999)

[Publications] Saito,I., et al.: "Fas ligand-mrdiated exocrinopathy resembling Sjogren's syndrome in mice transgenic for IL-10."J.Immunol.. 162. 2488-2494 (1999)

[Publications] Ikeda,A., et al.: "P300/CBP-dependent and-independent transcriptional interference between NK-kB RelA and p53."Biochem.Biophys.Res.Communs.. 272. 375-379 (2000)

[Publications] Ito,M., et al.: "Isoforms of JSAP1 scaffold protein generated through alternative splicing."Gene. 255. 229-234 (2000)

[Publications] S-W.Kee, et al.: "Human Hepatitis B Virus X protein is a possible mediator of hypoxia induced angiogenesis in hepatocellular carcinoma."Biochem.Res.Communs.. 268. 456-461 (2000)

[Publications] Dorjsuren,D.: "B型肝炎ウイルス(HBV)Xタンパクに拮抗する新規タンパクRMP。"実験医学. 18. 1137-1141 (2000)

[Publications] K.Masutomi, et. al,.: "Teromerase Activity Reconstituted in Vitro with Purified Human Teromerase Reverse Transcriptase and Human Teromerase RNA Component."J.Biol.Chem.. 275. 22568-22573 (2000)

[Publications] 増富健吉: "昆虫細胞発現系を用いたヒトテロメアーゼ逆転写酵素(hTERT)の発現・精製。"バイオインダストリー. 17. 35-42 (2000)

[Publications] Wei,W., et al.: "Direct interaction between the subunit RAP30 of TFHF and RNA polymeras subunit5 (RPB5) which contributes to the association between TFHF and RNA polymerase II."J.Biol.Chem. (in press). (2001)

[Publications] Kuboki,Y., et al.: "A scaffold protein in the c-Jun NH2-terminal kinase signaling pathways suppresses the extracelller signal-regulated kinase signaling pathway."J.Biol.Chem.. (in press). (2001)

[Publications] Qin,W., et al.: "Mutational Analysisof the Structure and Functions of Hepatitis C Virus RNA-dependent RNA polymerase."Hepatol.. (in press). (2001)

[Publications] 村上清史: "ヒトテロメアーゼ逆転写酵素(hTERT)の発現・精製とヒトテロメアーゼ活性の試験管内の再構成。"教育通信. (印刷中). (2001)

[Publications] 村上清史: "ウイルス肝炎-最新情報「B型肝炎ウイルス(HVB)の遺伝子制御」"富士レビオ株式会社. 271 (1999)

村上清史金沢大学, がん研究所, 教授 (90019878)