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Studies on a unique rRNA sequence found in the highly conserved functional region of silkworm ribosomes

Research Project

Project/Area Number 12660053
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 蚕糸・昆虫利用学
Research InstitutionShinshu University

Principal Investigator

UCHIUMI Toshio  Shinshu University, Faculty of Textile Science and Technology, Associate Professor, 繊維学部, 助教授 (50143764)

Co-Investigator(Kenkyū-buntansha) NAKAGAKI Masao  Shinshu University, Faculty of Textile Science and Technology, Professor, 繊維学部, 教授 (70135169)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2000: ¥1,200,000 (Direct Cost: ¥1,200,000)
Keywordsribosome / 28S rRNA / silkworm / ribosomal protein / GTPase center / site-directed mutagenesis / covariant base change / protein biosynthesis / 28SrRNA / 部位特異的塩基異変 / 翻訳装置 / リボソームRNA / GTPaseドメイン / 鱗翅類昆虫 / RNA-蛋白質相互作用 / P蛋白質
Research Abstract

The GTPase-associated domain in 23/28S rRNA is one of the most highly conserved functional regions throughout all organisms. We detected a unique nucleotide sequence within the GTPase-associated domain in silkworm species, in which the bases at positions 1094 and 1098 (numbering from Escherichia coli 23S rRNA) are C and G instead of the otherwise universally conserved bases U and A, respectively. These changes were also observed in four other moths, but not in organisms other than the moths. The RNA fragment covering the silkworm GTPase-associated domain showed two smearing bands in native gel electrophoresis in the absence of ribosomal proteins. However, these bands shifted to a single clear band, wheh silkworm ribosomal proteins P0, P1, P2, and eL12 bound to the RNA, suggesting that the GTPase-associated domain of silkworm 28S rRNA has a labile structural feature that is stabilized by binding of the ribosomal proteins. In order to know the functional significance of the covariant bas … More e change in the silkworm rRNA, we attempted the introduction of C-1094 and G-1098 into E. coli ribosomes by using a plasmid carrying E. coli rDNA (rrnB). So far, we detected the mutant rRNA in isolated ribosomes, suggesting that the mutant ribosomes were assembled in vivo. Further analyses of function of the mutant ribosomes were remained.
We also investigated interactions among ribosomal proteins P0, P1, and P2 by native gel electrophoresis. Mixing of P1 and P2 formed the P1-P2 heterodimer. This heterodimer, but neither P1 nor P2 alone, tightly bound to P0 and formed a pentameric complex P0(P1-P2)_2, We confirmed the functions of the ribosomal proteins by incorporating the proteins into E. coli ribosomes, i.e., by replacing E. coli proteins L10-L7/L12 complex and L11 on the GTPase-associated RNA domain with silkworm P0(P1-P2)_2 complex and eL12. The protein replacement caused ribosomal activity with silkworm translation factors. This protein-dependent induction of the ribosomal activity did not occur in the absence either of P1 or P2. These results suggest that formation of P1-P2 heterodimer is crucial for assembly of the proteins on the rRNA domain and for the stabilization of the functional rRNA structure. Less

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Uchiumi, T. et al.: "A covariant change of the two highly conserved bases in the GTPase-associated center of 28 S rRNA in silkworms and other moths"J. Biol. Chem. 275. 35116-35121 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 内海利男: "リボソーム機能部位が見えてきた:X線結晶解析が示す翻訳装置の構造"蛋白質・核酸・酵素. 45. 1237-1244 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Anderson, C.J. et al.: "Autoantibodies to the 20-kDa ribosomal proteins : Identification, characterization, and new aspects on prevalence in systemic lupus erythematosus"Clinical Immunology. 98. 249-257 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Uchiumi, T. et al.: "Translation elongation by a hybrid ribosome in which proteins at the GTPase center of the Escherichia coli ribosome are replaced with rat counterparts"J. Biol. Chem.. 277. 3857-3862 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shimizu, T. et al.: "Interaction among silkworm ribosomal proteins P1,P2 and P0 required for functional protein binding to the GTPase-associated domain of 28S rRNA"Nucleic Acids Res.. 30. 2620-2627 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Uchiumi, T. et al.: "Ribosomal proteins at the stalk region modulate functional rRNA structures in the GTPase center"J. Biol. Chem. 277. 41401-41409 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 内海利男: "遺伝子発現研究法(生物化学実験法43);江尻慎一郎, 平秀晴, 堤賢一, 志村憲助 編集;「IX.リボソーム」"学会出版センター. 143-159 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Uchiumi, T., Nomura, T., Shimizu, T., Katakai, Y., Mita, K., Koike, Y., Nakagaki, M., Taira, H. and Hachimori, A.: "A covariant change of the two highly conserved bases in the GTPase-associated center of 28 S rRNA in silkworms and other moths."J. Biol. Chem.. 275. 35116-35121 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Uchiumi, T.: "Functional structures in the ribosome revealed by a crystallographic analysis."Tanpakushitsu Kakusan Koso. 45. 1237-1244 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Anderson, C. J., Neas, B. R., Uchiumi. T. and Stafford, H. A.: "Autoantibodies to the 20-kDa ribosomal proteins: Identification, characterization, and new aspects on prevalence in systemic lupus erythematosus."Clinical Immunology. 98. 249-257 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Uchiumi T., Honma, S., Nomura, T., Dabbs, E. R. and Hachimori, A.: "Translation elongation by a hybrid ribosome in which proteins at the GTPase center of the Escherichia coli ribosome are replaced with rat counterparts."J. Biol. Chem.. 277. 3857-3862 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shimizu, T., Nakagaki, M., Nishi, Y., Kobayashi, Y., Hachimori, A., and Uchiumi, T.: "Interaction among silkworm ribosomal proteins P1, P2 and P0 required for functional protein binding to the GTPase-associated domain of 28S rRNA."Nucleic Acids Res.. 30. 2620-2627 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Uchiumi, T., Honma, S., Endo, Y., and Hachimori, A.: "Ribosomal proteins at the stalk region modulate functional rRNA structures in the GTPase center."J. Biol Chem.. 277. 41401-41409 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shimizu, T., Uchiumi T.et al.: "Interaction among silkworm ribosomal proteins P1, P2 and PO required for functional protein binding to the GTPase-associated domain of 28S rRNA"Nucleic Acids Res.. 30. 2620-2627 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Uchiumi, T.et al.: "Translation elongation by a hybrid ribosome in which proteins at the GTPase center of the Escherichia coli ribosome are replaced with rat counterparts"J. Biol. Chem.. 277. 3857-3862 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Uchiumi T.et al.: "Ribosomal proteins at the stalk region modulate functional rRNA structures in the GTPase center"J. Biol. Chem.. 277. 41401-41409 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hasegawa, H., Uchiumi, T.et al.: "Autoantibody against ribosomal protein L14 in patients with systemic lupus erythematosus"Clin. Exp. Rheumatol.. 20. 139-144 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Uchiumi, T., et al.: "Translation elongation by a hybrid ribosome in which proteins at the GTPase center of the Escherichia coli ribosome are replaced with rat counterparts"J. Biol. Chem.. 277. 3857-3862 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Shizawa, N., et al.: "Direct mutagenesis studies of the C-terminal fingerprint region of Bacillus subtilis Pyrophosphatase"Eur. J. Biochem.. 268. 5771-5775 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Uchiumi,T. et al.: "A Covariant Change of the Two Highly Conserved Bases in the GTPase-associated Center of 28 S rRNA in Silkworms and Other Moths."J.Biol.Chem.. 275. 35116-35121 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Anderson,C.J. et al.: "Autoantibodies to the 20-kDa Ribosomal Proteins : Identification, Characterization, and New Aspects on Prevalence in Systemic Lupus Erythematosus."Clinical Immunol.. 98. 249-253 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 内海利男: "リボソームの機能部位が見えてきた:X線結晶解析が示す翻訳装置の構造."蛋白質・核酸・酵素. 45. 1237-1244 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 石川冬木 他: "RNA研究の最前線"シュプリンガー・フェアラーク東京. 238 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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