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Molecular mechanism and physiological roles of trans-translation mediated by tmRNA

Research Project

Project/Area Number 14380322
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Molecular biology
Research InstitutionHirosaki University

Principal Investigator

MUTO Akira  Hirosaki University, Faculty of Agriculture and Life Science, Professor, 農学生命科学部, 教授 (80034635)

Co-Investigator(Kenkyū-buntansha) HIMENO Hyouta  Hirosaki University, Faculty of Agriculture and Life Science, Associate Professor, 農学生命科学部, 助教授 (80208785)
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥14,200,000 (Direct Cost: ¥14,200,000)
Fiscal Year 2004: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2003: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2002: ¥7,800,000 (Direct Cost: ¥7,800,000)
KeywordstmRNA / tran-tranalation / Escherichia coli / Bacillus subtilis / antibiotics / ribosome / tag-peptide / spore / RNA / 翻訳 / 遺伝子 / SmpB
Research Abstract

a)Molecular mechanism of trans-translation mediated by tmRNA
To investigate the functional structure of tmRNA in trans-translation reaction, we have developed an in vitro trans-translation system using E.coli extracts. Many mutant tmRNAs were constructed by in vitro mutagenesis, and tested their activities as tRNA, mRNA and rribosome binding. From the results, we could identify the structures and sequences in tmRNA required for each function. The effect of several antibiotics on the trans-translation was investigated : we found that paromomycin directly binds to tmRNA causing the frame-shift of translation of tab peptides.
b)Physiological roles of trans-translation
We have constructed an in vivo system in which trans-translation products can be detected in Bacillus subtilis and E.coli. Using this system, several reaction products were isolated and identified by the N-terminal amino acid sequence analyzes. The sites of trans-translation in some of the products were determined by MAS-spectrometry. It was shown that the trans-translation is involved in various cellular activities, such as catabolite repression, amino acid metabolism, stress responses, sporulation and so on.
Endspore formation of Bacillus subtilis strain is significantly impaired by deleting the gene for tmRNA. The genetic and biochemical experimental results revealed that the synthesis of sigma-Kduring the late stage of sporulation process is significantly impaired in the mutant cells.

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (22 results)

All 2005 2004 Other

All Journal Article (12 results) Publications (10 results)

  • [Journal Article] Participation of 3'-to-5' Exoribonucleases in the turnover of Bacillus subtilis Messenger RNA2005

    • Author(s)
      Oussenko, I., Abe, T., Ujiie, H., Muto, A., Bechhofer, D.A.
    • Journal Title

      J.Bacteriology 187

      Pages: 2758-2767

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Participation of 3'-to-5' Exoribonucleases in the turnover of Bacillus subtilis Messenger RNA2005

    • Author(s)
      Oussenko, I., Abe, T., Ujiie, H., Muto, A., Bechhofer, D.A.
    • Journal Title

      J.Bacteriol. 187

      Pages: 2758-2767

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Participation of 3'-to-5' Exoribonucleases in the turnover of Bacillus subtilis Messenger RNA2005

    • Author(s)
      Oussenko, I., Abe, T., Ujiie, H., Muto, A., Bechhofer, D.A.
    • Journal Title

      J.Bacteriology 187(in press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A minimum structure of aminoglycosides that causes an initiation shift of trans-translation2004

    • Author(s)
      Konno, T., Takahashi, T., Kurita, D., Muto, A., Himeno, H
    • Journal Title

      Nucleic Acids Res. 32

      Pages: 4119-4126

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Isolation of eight novel Caenorhabditis elegans small RNAs2004

    • Author(s)
      Wachi, M., Ogawa, T.Yokoyama, K.Hokii, Y., Shimoyama, M., Muto, A., Ushida, C.
    • Journal Title

      Gene 335

      Pages: 47-56

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A novel GTPase activated by small subunit of ribosome2004

    • Author(s)
      Himeno, H., Hanawa-Suetsugu, K., Kimura, T., Takagi, K., Sugiyama, W., Shirata, S., Mikami, T., Odagiri, F., Osanai, Y., Watanabe, D., Goto, S., Kalachnyuk, L., Ushida, C., Muto, A.
    • Journal Title

      Nucleic Acids Res. 32

      Pages: 5303-5309

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] 翻訳と共役した蛋白質品質管理システム:トランストランスレーション2004

    • Author(s)
      姫野俵太, 武藤 あきら
    • Journal Title

      蛋白質核酸酵素 49

      Pages: 1062-1066

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Isolation of eight novel Caenorhabditis elegans small RNAs2004

    • Author(s)
      Wachi, M., Ogawa, T., Yokoyama, K., Hokii, Y., Shimoyama, M., Muto, A., Ushida, C.
    • Journal Title

      Gene 335

      Pages: 47-56

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A novel GTPase activated by the small subunit of ribosome2004

    • Author(s)
      Himeno, H., Hanawa-Suetsugu, K., Kimura, T., Takagi, K., Sugiyama, W., Shirata, S., Mikami, T., Odagiri, F., Osanai, Y., Watanabe, D., Goto, S., Kalachnyuk, L., Ushida, C., Muto, A.
    • Journal Title

      Nucelic Acids Res. 32

      Pages: 5303-5309

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Trans-tranlation : Co-translational protein quality control system2004

    • Author(s)
      Himeno, H., Muto, A.
    • Journal Title

      Protein, Nucleic Acid and Enzyme 49(7)(suppl.)

      Pages: 1062-1066

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Isolation of eight novel Caenorhabditis elegans small RNAs2004

    • Author(s)
      Wachi, M., Ogawa, T., Yokoyama, K, Hokii, Y., Shimoyama, M., Muto, A., Ushida, C.
    • Journal Title

      Gene 335

      Pages: 47-56

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A novel GTPase activated by the small subunit of ribosome2004

    • Author(s)
      Himeno, H., Hanawa-Suetsugu, K., Kimura, T., Takagi, K., Sugiyama, W., Shirata, S., Mikami, T., Odagiri, F., Osanai, Y., Watanabe, D., Goto, S., Kalachnyuk, L., Ushida, C., Muto, A.
    • Journal Title

      Nucleic Acids Res. 32

      Pages: 5303-5309

    • Related Report
      2004 Annual Research Report
  • [Publications] T.Takahashi, T.Konno, A.Muto, H.Himeno: "Various effects of paromomycin on tmRNA-mediated trans-translation"J.Biol.Chem.. 278. 27672-27680 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] C.Ushida, A.Yoshida, Y.Miyakawa, Y.Ara, A.Muto: "Distribution of the MCS4 RNA genes in mycoplasmas belonging to the Mycoplasma mycoides cluster"Gene. 314. 149-155 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 武藤あきら, 姫野俵太: "tmRNAによるトランス翻訳機構と生理的機能"蛋白質核酸酵素. 48. 346-356 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.Konno, T.Takahashi, A.Muto, H.Himeno: "Various effect of paramomycin on tmRNA-mediated trans-translation"Nucleic Acid Res. Supplement. 3. 235-236 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] K.Takada, C.Hori-takemoto, M.Kawazoe, S.Lee, M.Shirouzu, M.Yakoyama, A.Muto, H.Himeno: "In vitro anaysis of the initiation steps of trans-translation"Nucleic Acid Res. Supplement. 3. 235-236 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 姫野 俵太: "タンパク質がわかる"竹縄忠臣編(羊土社). 26-35 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] A.Fujihara, H.Tomatsu, S.Inagaki, C.Ushida, H.Himeno, A.Muto: "Detection of tmRNA-mediated trans-translation products in Bacillus subtilis"Genes to Cells. 7. 343-350 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Hanawa-Suetsugu, H.Himeno, A.Muto: "SmpB functions in various steps of trans-translation"Nucelic Acids Research. 30. 1620-1629 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Ito, T.Tadaki, S.Lee, K.Takada, A.Muto, H.Himeno: "Trans-translation mediated by Bacillus subtilis tmRNA"FEBS Letters. 516. 245-252 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 渡辺公綱, 姫野俵太: "生命科学II 遺伝子の働きとその応用"丸善株式会社. 255 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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