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2002 Fiscal Year Final Research Report Summary

Mechanism of transcription of Paramyxovirus genome : Purification and characterization of host factors

Research Project

Project/Area Number 11470080
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Virology
Research InstitutionKitasato University

Principal Investigator

MIZUMOTO Kiyohisa  Kitasato University, School of Pharmaceutical Sciences, Professor, 薬学部, 教授 (80092344)

Co-Investigator(Kenkyū-buntansha) OGINO Tomoaki  University of Tokyo, Graduate School Researcher, 医学系研究科, 研究員 (80312213)
IWAMA Minako  Kitasato University, School of Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (10223421)
TSUKAMOTO Toshihiko  Kitasato University, School of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (10236862)
Project Period (FY) 1999 – 2002
KeywordsParamyxovirus / Sendai virus / negative-strand RNA genome / RNA-dependent RNA polymerase / host factor / transcription / mRNA cap / tubulin
Research Abstract

Sendai virus (SeV), a member of Paramyxovirus family, contains a monopartite negative strand RNA genome of approximately 15 kilobases. To elucidate the mechanism of transcription and replication of SeV, we developed an efficient and faithful in vitro transcription system using purified virus particles. We have previously shown that in vitro mRNA synthesis of SeV is almost entirely dependent on the addition of cellular proteins (host factors). In the present study, we further furified and characterized the host factor activity, and obtained the following results. 1) We purified the host factor activity, which stimulates in vitro SeV mRNA synthesis, from bovine brain, and found that the activity consists of four complementary protein factors, tubulin, phosphoglycerate kinase, enolase, and unidentified p24. 2) We showed that tubulin is involved in a transcription initiation complex formation by removing the viral M protein from the RNP, and by being integrated itself into the complex. On the other hand, phosphoglycerate kinase, enolase, and p24 are found to act at RNA shain elongation step. Furthermore, 3) we demonstrated that a recombinant viral L protein exhibits the cap methylase (mRNA guanine-7-methyltransferase) activity.

  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] K.Masunaga et al.: "Molecular mapping of influenza virus RNA polymerase by site-specific antibodies"Virology. 256. 130-141 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S.Muraoka et al.: "Detection and identification of amanitins in the wood-rotting fungi Galerina fasciculata and Galerina helvoliceps"Appl. Env. Microbiol.. 65. 4207-4210 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Honda et al.: "Two separate sequences of PB2 subunit constitute the RNA cap-binding site of influenza virus RNA polymerase"Genes to Cells. 4. 475-485 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Ogino et al.: "Involvement of a cellular glycolytic enzyme, phosphoglycerate kinase, in Sendai virus transcription"J. Biol. Chem.. 274. 35999-36008 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Yokoska et al.: "Cloning and characterization of mRNA capping enzyme and mRNA(guanine-7-)methyltransferase cDNAs from Xenopus laevis"Biochem. Biophys. Res. Commun.. 268. 617-624 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] E.Kajita et al.: "Isolation and characterization of Xenopus laevis aldolase B cDNA and expression patterns of aldolase A, B, and C genes in adult tissues, oocytes and embryos of Xenopus laevis"Biochem. Biophys. Acta. 1493. 101-118 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Chiba et al.: "Actinohivin, a novel anti-HIV protein from an actinomycete that inhibits syncytium formation : isolation characterization, and biological activities"Biochem. Biophys. Res. Commun.. 282. 595-601 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Ogino et al.: "Enolase, a cellular glycolytic enzyme, is required for ellicient transcriptio of Sendai virus genome"Biochem. Biophys. Res. Commun.. 285. 447-455 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Honda et al.: "Differential roles of viral RNA and cRNA in functional modulation of the influenza virus RNA polymerase"J. Biol. Chem.. 276. 31179-31185 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Ymaochi et al.: "ik3-1/Cables is a substrate for cyclin-dependent kinase 3 (cdk 3)"Eur. J. Biochem.. 268. 6076-6082 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Tsuji et al.: "Differential ellect of ik3-1/Cables on p53-and p73-induced cell death"J. Biol. Chem.. 277. 2951-2957 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] N.Radomski et al.: "Interaction of S-adenosylhomocysteine hydrolase of Xenopus laevis with mRNA (guanine-7-)methyltransferase : Implication on its nuclear compartmentalisation and on cap methylation of hnRNA"Biochem. Biophys. Acta. 1590. 93-102 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Honda et al.: "Minimum molecular architectures for transcription and replication of the influenza virus"Proc. Natl. Acad. Sci. USA. 99. 13166-13171 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Ochi et al.: "Proteomic analysis of human brain identifies a-enolase as a novel autoantigen in Hashimoto's encephalopathy"FEBS Letters. 528. 197-202 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Tsuji et al.: "p53-independent apoptosis is induced by the p19ARF tumor suppressor"Biochem. Biophys. Res. Commun.. 295. 621-629 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Masunaga, K.Mizumoto, H.Kato, A.Ishihama, T.Toyoda: "Molecular mapping of influenza virus RNA polymerase by site-specific antibodies"Virology. 256. 130-141 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Muraoka, N.Fukamachi, K.Mizumoto, T.Shinozawa: "Detection and identification of amanitins in the wood-rotting fungi Galerina fasciculata and Galerina helvoliceps"Appl.Env.Microbiol.. 65. 4207-4210 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.Honda, K.Mizumoto, and A.Ishihama: "Two separate sequences of PB2 subunit constitute the RNA cap-binding site of influenza virus RNA polymerase"Genes to Cells. 4. 475-485 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Ogino, M.Iwama, J.Kinouchi, Y.Shibagaki, T.Tsukamoto, K.Mizumoto: "Involvement of a cellular glycolytic enzyme, phosphoglycerate kinase, in Sendai virus transcription"J.Biol.Chem.. 274. 35999-36008 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Yokoska, T.Tsukamoto, K.Miura, K.Shiokawa, K.Mizumoto: "Cloning and characterization of mRNA capping enzyme and mRNA (guanine-7-) methyltransferase cDNAs from Xenopus laevis"Biochem.Biophys.Res.Commun.. 268. 617-624 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] E.Kajita, M.Wakiyama, K.Miura, K.Mizumoto, T.Oka, I.Komuro, T.Miyata, H.Yatsuki, K.Hori, and K.Shiokawa: "Isolation and characterization of Xenopus laevis aldolase B cDNA and expression patterns of aldolase A, B, and C genes in adult tissues, oocytes and embryos of Xenopus laevis"Biochem.Biophys.Acta.. 1493. 101-118 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Chiba, J.Inokoshi, M.Okamoto, S.Asanuma, K.Matsuzaki, M.Iwama, K.Mizumoto, H.Tanaka, M.Oheda, K.Fujita, H.Nakashima, M.Shinose, Y.Takahashi, S.Omura: "Actinohivin, a novel anti-HIV protein from an actinomycete that inhibits syncytium formation : isolation icharacterization, and biological activities"Biochem.Biophys.Res.Commun.. 282. 595-601 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Ogino, T.Yamadera, T.Nonaka, S.Imajoh-Ohmi, K.Mizumoto: "Enolase, a cellular glycolytic enzyme, is required for ellicient transcriptio of Sendai virus genome"Biochem.Biophys.Res.Commun.. 285. 447-455 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.Honda, A.Endo, K.Mizumoto, and A.Ishihama: "Differential roles of viral RNA and cRNA in functional modulation of the influenza virus RNA polymerase"J.Biol.Chem.. 276. 31179-31185 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Yamochi, K.Tsuji, K.Mizumoto, H.Sato, K.Senba, Y.Matsuura, I.Nishimoto, M.Matsuoka: "ik3-1/Cables is a substrate for cyclin-dependent kinase 3 (cdk 3)"Eur.J.Biochem.. 268. 6076-6082 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Tsuji, K.Mizumoto, T.Yamochi, I.Nishimoto, M.Matsuoka: "Differential ellect of ik3-1/Cables on p53- and p73-induced cell death"J.Biol.Chem.. 277. 2951-2957 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] N.Radomski, G.Barreto, C.Kaufmann, J.Yokoshuka, K.Mizumoto, C.Dreyer: "Interaction of S-adenosylhomocysteine hydrolase of Xenopus laevis with mRNA(guanine-7-)methyltransferase : Implication on its nuclear compartmentalisation and on cap methylation of hnRNA"Biochem.Biophys.Acta.. 1590. 93-102 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.Honda, K.Mizumoto, A.Ishihama: "Minimum molecular architectures for transcription and replication of the influenza virus"Proc.Natl.Acad.Sci.USA. 99. 13166-13171 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Ochi, I.Horiuchi, N.Araki, T.Toda, K.Sato, H.Murai, M.Osoegawa, T.Yamada, K.Okamura, T.Ogino, K.Mizumoto, H.Yamashita, H.Saya, J.Kira: "Proteomic analysis of human brain identifies a-enolase as a novel autoantigen in Hashimoto's encephalopathy"FEBS Letters. 528. 197-202 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Tsuji, K.Mizumoto, H.Sudo, K.Kouyama, E.Ogata, and M.Matsuoka: "p53-independent apoptosis is induced by the p19ARF tumor suppressor"Biochem.Biophys.Res.Commun.. 295. 621-629 (2002)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2004-04-14  

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