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2000 Fiscal Year Final Research Report Summary

Structural and functional alayses of the plant viral RNA polymerases for the breeding of virus-resistant transgenic plants.

Research Project

Project/Area Number 10356002
Research Category

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

Allocation TypeSingle-year Grants
Section展開研究
Research Field 植物保護
Research InstitutionTHE UNIVERSITY OF TOKYO

Principal Investigator

HIBI Tadaaki  U.Tokyo, Grad S., Agr.Life Sci., Prof., 大学院・農学生命科学研究科, 教授 (50261954)

Co-Investigator(Kenkyū-buntansha) OOHIRA Kazuyuki  Suntory Co., Basic Inst., Chief Researcher, 基礎研究所, 主任研究員
SUZUKI Masasi  U.Tokyo, Grad S., Agr.Life Sci., Assist.Prof., 大学院・農学生命科学研究科, 助手 (40282694)
SHIRAKO Yukio  U.Tokyo, Asian Nat.Environ.Sci.Center, Prof., アジア生物資源環境研究センター, 教授 (90143023)
Project Period (FY) 1998 – 2000
Keywordsplant virus / RNA polymerase / host protein / TMV / CMV / SBWMV / pseudorecombinant / virus-resistant transgenic plant
Research Abstract

1. Active and template-specific tobacco mosaic virus (TMV) RNA polymerase was successfully purified from TMV-infected tobacco leaf extracts through an affinity chlomatography using anti-RNA polymerase domein of TMV 183K protein. The apoenzyme was presumed to be a heterodimer composed of each one molecules of 126K and 183K proteins. While, three different host proteins bound to helicase domein of TMV RNA polymerase were detected by the yeast two-hybrid system.
2. The genome of both cucumber mosaic virus (CMV) and peanut stunt virus (PSV) consists of three single-stranded RNAs, RNA 1, 2 and 3. 1a protein and 2a protein encoded by RNA1 and RNA2, respectively, are necessary for the viral replication. Feasibility of exchanges of RNAs 1 and 2 was re-examined by using infectious RNA transcripts for CMV RNAs and PSV RNAs. When we coinoculated all the pseudorecombinants into tobacco protoplasts, pseodorecombinants with PSV RNA 1 and CMV RNA 2 were replicable, but those with CMV RNA 1 and PSV RNA … More 2 were not. Using yeast two-hybrid system, the C terminal region of PSV-1a protein was exhibited to interact with not only the N terminal region of PSV 2a protein but also that of CMV 2a protein. These results were correlated with those of replication in protoplasts.
3. Using full-length infectious cDNA clones to RNA 1 and RNA2 of Soil-borne wheat mosaic virus (SBWMV), a series of RNA 1 mutants of 152K and 211K replicase proteins were constructed. Results showed that the 211K protein with methyltransferase, helicase and polymerase domains in the absence of the 152K protein possessing the former two domains is sufficient for virus replication at a low level. The 152K protein together with the 211K protein were shown to be required for the high-level RNA replication. Expression of a cysteine-rich protein encoded in the 3'-terminal region of RNA2 was absolutely required for virus replication. Infectious cDNA clones to a US strain were constructed. Reassortment experiments between US and Japanese strains showed that the combination of Japanese RNA1 and US RNA2 was systemically infectious to wheat plants whereas that of US RNA1 and Japanese RNA2 was not infectious even to Chenopodium quinoa, a local lesion host. Less

  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Masuta,C.,Ueda,S.,Suzuki,M.and Uyeda,I.: "Evolution of a novel quadripartite hybrid virus by interspecific exchange and recombination between replicase components of two related tripartite RNA viruses."Proceedings of National Academy of Sciences of USA. 95. 10487-10492 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shirako,Y.: "Non-AUG translation initiation in a plant RNA virus genome : a forty-amino-acid extension is added to the N-terminus of the soil-borne wheat mosaic virus capsid protein."J.Virology. 72. 1677-1682 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Watanabe,T.,Honda,A.,Iwata,A.,Ueda,S.,Hibi,T. et al.: "Isolation from tobacco mosaic virus-infected tobacco of a solubilized template-specific RNA-dependent RNA polymerase containing a 126K/183K protein heterodimer."J.Virology. 73. 2633-2640 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takemoto,Y.,Kanehira,T.,Shinohara,M.,Hibi,T. et al.: "The nucleotide sequence and genomic organization of Japanese iris necrotic ring virus, a new species in the genus Carmovirus."Archives of Virology. 145. 651-657 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miranda,G.J.,Azzam,O.,and Shirako,Y.: "Comparison of nucleotide sequences between northern and southern Philippine isolates of rice grassy stunt virus indicates occurrence of natural genetic reassortment."Virology. 266. 26-32 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shirako,Y.,Suzuki,N.,and French.R.C.: "Similarity and divergence among viruses in the genus Furovirus."Virology. 270. 201-207 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamiya,A.,and Shirako,Y.: "Construction of full-length cDNA clones to Soil-borne wheat mosaic virus RNA1 and RNA2 from which infections RNAs are transcribed in vitro."Virology. 277. 66-75 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miranda,G.J.,Aliyari,R.,and Shirako,Y.: "Nucleotide sequence of a Dianthovirus RNA1-like RNA found in grassy stunt-diseased rice plants."Archives of Virology. 145. 1-14 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 渡辺貴斗,日比忠明: "植物プラス1本鎖RNAウイルスのRNA複製酵素."ウイルス. 50. 103-118 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takemoto,Y.,Hibi,T.: "Soybean chlorotic mottle virus genes Ia, II, III, IV and V are essential but Ib product is nonessential for the systemic infection."J.General Virology,.. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takahashi,H.,Suzuki,M., et al.: "Mapping of virus and host genes which confer the resistance response in Cucumber mosaic virus-infected Arabidopsis thaliana."Plant and Cell Physiology. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takeshita,M.,Suzuki,M.and Takanami Y.: "Combination of amino acids in the 3a protein and the coat protein of cucumber mosaic virus determines symptom expression and viral spread in bottle gourd."Archives of Virology. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masuta, C., Ueda, S., Suzuki, M.and Uyeda, I.: "Evolution of a novel quadripartite hybrid virus by interspecific exchange and recombination betweenreplicase components of two related tripartite RNA viruses."Proc.Natl.Acad.Sci.USA. 95. 10487-10492 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shirako, Y.: "Non-AUG translation initiation in a plant RNA virus genome : a forty-amino-acid extension is added to the N-terminus of the soil-borne wheat mosaic virus capsid protein."J.Virol.. 72. 1677-1682 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe, T., Honda, A., Iwata, A., Ueda, S., Hibi, T.et al.: "Isolation from tobacco mosaic virus-infected tobacco of a solubilized template-specific RNA-dependent RNA polymerase containing a 126K/183K protein heterodimer."J.Virol.. 73. 2633-2640 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takemoto, Y., Kanehira, T., Shinohara, M., Hibi, T.et al.: "The nucleotide sequence and genomic organization of Japanese iris necrotic ring virus, a new species in the genus Carmovirus."Arch Virol.. 145. 651-657 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miranda, G.J., Azzam, O., and Shirako, Y.: "Comparison of nucleotide sequences between northern and southern Philippine isolates of rice grassy stunt virus indicates occurrence of natural genetic reassortment."Virology. 266. 26-32 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shirako, Y., Suzuki, N., and French, R.C.: "Similarity and divergence among viruses in the genus Furovirus."Virology. 270. 201-207 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamiya, A., and Shirako, Y.: "Construction of full-length cDNA clones to Soil-borne wheat mosaic virus RNA1 and RNA2, from which infectious RNAs are transcribed in vitro."Virology. 277. 66-75 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miranda, G.J., Aliyari, R., and Shirako, Y.: "Nucleotide sequence of a Dianthovirus RNA1-like RNA found in grassy stunt-diseased rice plants."Arch.Virol.. 145. 1-14 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takemoto, Y., Hibi, T.: "Soybean chlorotic mottle virus genes Ia, II, III, IV and V are essential but Ib product is nonessential for the systemic infection."J.Gen.Virol.. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takahashi, H., Suzuki, M.et al.: "Mapping of virus and host genes which confer the resistance response in Cucumber mosaic virus-infected Arabidopsis thaliana."Plant and Cell Physiol.. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takeshita, M., Suzuki, M.and Takanami Y.: "Combination of amino acids in the 3a protein and the coat protein of cucumber mosaic virus determines symptomexpression and viral spread in bottle gourd."Arch.Virol.. (in press). (2001)

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

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Published: 2002-03-26  

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