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Analysis on mechanisms of brome mosaic virus RNA replication

Research Project

Project/Area Number 04404009
Research Category

Grant-in-Aid for General Scientific Research (A)

Allocation TypeSingle-year Grants
Research Field 植物保護
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

FURUSAWA Iwao  KYOTO UNIV.FAC.AGRI.PROFESSOR, 農学部, 教授 (10026594)

Co-Investigator(Kenkyū-buntansha) YUBO Yasuyuki  KYOTO PRIFECTURAL UNIV.FAC.AGRI.ASSISUTANT PROFESSOR, 農学部, 助教授 (80183797)
OKUNO Teturou  KYOTO UNIV.FAC.AGRI.ASSISUTANT PROFESSOR, 農学部, 助教授 (00221151)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥20,500,000 (Direct Cost: ¥20,500,000)
Fiscal Year 1994: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1993: ¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1992: ¥12,000,000 (Direct Cost: ¥12,000,000)
Keywordsbrome mosaic virus / monoclonal antibody / RNA replicase / immunoelectron microscopy / epitope mapping / BMV / 形質転換植物 / 試験管内転換 / 試験管内翻訳 / トランスジェニック植物 / RNA複製酵素遺伝子 / 非宿主植物 / トランス / ゲル固定化酵素
Research Abstract

Cellular structures or micro-organella involved in virus RNA replication have been debated. RNA dependent RNA polymerase (RdRp) specific to virus infection is extracted from BMV infected barley plants. The RdRp was associated with membrane structure and RNA templates in the infected cells. Ultrastructural study of the BMV-infection specific membrane structure using BMV RNA specific probe has not been successful to assign more detailed structure for sites of virus RNA replication. Now, BMV replicase components, 1a and 2a proteins produced in E.coli are available to obtain monoclonal antibody which can be used to make epitope mapping of the proteins. Immunoelectron microscopic study of BMV-infected cells using monoclonal antibody against BMV 1a protein indicated that mebrane structures specifically developed to virus infection and the structures contained 1a protein specific gold particles. The membrane structures were distinct from micro-organella usually observed in healthy barley cells. The infection specific structures developed with infection stage. Similar study using membrane fraction of BMV infected barley plants showed that gold particles specific to BMV 1a protein was not associated with any micro-organella present in healthy cells. Purified BMV RdRp fraction was used for similar study. When RdRp fraction treated with ionic detergent (SDS) was fixed on a mesh and reacted with 1a monoclonal antibody, 1a protein specific gold particles were observed in single or double, while gold particles were observed in a cluster containing 4-7 particles in samples untreated with SDS.This suggests that BMV 1a protein exists as oligomer and that BMV replicase may consist of the 1a oligomer. Epitope mapping of BMV 1a protein was made using 11 monoclonal anitodies against 1a protein.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Mori,M., Mise.K., Okuno,T.and Furusawa,I.: "Expression of brome mosaic virus-encoded replicase genes in transgenic tobacco plants." J.gen.Virol.73. 169-172 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mori,M.,Zhang,G-H.,Okuno,T.and Furusawa,I.: "Efficient production of human gamma interferon in tobacco protoplasts by genetically engineered brome mosaic virus RNAs." J.gen.Virol.74. 1255-1260 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mori,M.,Kaido,M.,Okuno,T.and Furusawa,I.: "mRNA amplification system by viral replicase in transgenic plants." FEBS Lett. 336. 171-174 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mori, M., Mise, K., Okuno, T.and Furusawa, I.: "Expression of brome mosaic virus-encoded replicase genes in transgenic tobacco Plants." J.gen.Virol.73. 169-172 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mori, M., Zhang, G-H., Okuno, T.and Furusawa, I.: "Efficient production of human gamma interferon in tobacco protoplasts by genetically engineered brome mosaic virus RNAs." J.gen.Virol.74. 1255-1260 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mori, M., Kaido, M., Okuno, T.and Furusawa, I.: "mRNA amplification system by viral replicase in transgenic plants." FEBS Lett. 336. 171-174 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mori,M.,Mise,K.,Okuno,T.and Furusawa,I.: "Expression of brome mosaic virus-encoded replicase genes in transgenic tobacco plants." J.gen.Virol.73. 169-172 (1992)

    • Related Report
      1994 Annual Research Report
  • [Publications] Mori,M.,Zhang,G-H.,Okuno,T.and Furusawa,I.: "Efficient production of human gamma interferon in tobacco protoplasts by genetically engineered brome mosaic virus RNAs." J.gen.Virol.74. 1255-1260 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] Mori,M.,Kaido,m.,Okuno,T.and Furusawa,I.: "mRNA amplification system by viral replicase in transgenic plants." FEBS Lett. 336. 171-174 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Mori,M.Kaido,T.Okuno,and I.Furusawa: "mRNA amplification system by viral replicase in transgenic plants" FEBS LETTERS. 336. 171-174 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Mori,G.Zhang,M.Kaido,T.Okuno,and I.Furusawa: "Efficient production of human gamma interferon in tobacco protoplasts by genetically engineered rome mosaic virus RNAs" Journal of General Virology. 74. 1255-1260 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Mori,Mise,T.Okuno and I.Furusawa: "Expression of brome mosaic virus-encoded replicase genes in transgenic tobacco plants" Journal of Geneval Virology. 73. 169-172 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Mise,S.Tsuge,K.Nagao,T.Okuno and I.Furusawa: "Nucleotide sequence responsible for the synthesis of a truncated coat protein of brome mosaic virus strain ATCC66" Journal of General Virology. 73. 2543-2551 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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