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Molecular mechanism of the animal mitochondrial translation systems

Research Project

Project/Area Number 08408025
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionThe University of Tokyo

Principal Investigator

WATANABE Kimitsuna  The University of Tokyo, Graduate School of Engineering, Prof., 大学院・工学系研究科, 教授 (00134502)

Co-Investigator(Kenkyū-buntansha) NITTA Itoru  The University of Tokyo, Graduate School of Engineering, Assistant Prof., 大学院・工学系研究科, 助手 (30272404)
KAWAI Gota  Chiba Institute of Technology, Faculty of Engineering, Assistant Prof., 工学部, 講師 (70211860)
UEDA Takuya  The University of Tokyo, Graduate School Frontier Science, Associate Prof., 大学院・新領域創成科学研究科, 助教授 (80184927)
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥8,900,000 (Direct Cost: ¥8,900,000)
Fiscal Year 1998: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1997: ¥5,400,000 (Direct Cost: ¥5,400,000)
Keywordsanimal mitochondria / in vitro translation system / non-universal genetic code / tRNA / modified base / AUA codon / aminoacyl-tRNA synthetase / elongation factors / 非普遍暗号 / Wobble則 / ホルミルトランスフェラーゼ / セリル-tRNAシンテターゼ / イン-ビトロ翻訳系 / AVAコドン / クローバ葉型構造
Research Abstract

This research aims to elucidate the molecular mechanism of animal mitochondrial (mt) translation system through carrying out (1) structural analysis of animal mt tRNAs, and (2) verification of translation ability of these tRNAs by using a mitochondrial in vitro translation system. The animal mt translation system possesses the distinct features as compared with those of prokaryotic and eukaryotic cytoplasmic systems ; exsistences of tRNA possessing unusual secondary structures and non-universal genetic code, only 22 species of tRNAs exist in mitochondria, a single tRNA^<Met> is used for both initiator and elongator, ribosomes contain rRNA with shorter chain lengths and much more proteins. We have isolated and characterized individual factors involved in the translation system and got the following new findings. 1) We found several non-universal genetic codes in various animal nit systems and analyzed the primary structures of tRNAs decoding the codes. it turned out that the genetic code variations are mainly induced by the base (mainly modified base) in the anticodon first position (Wobble position), which lead to the establishment of the mt specific Wobble rule. 2) By combining biochemical experiments and ^1H-NMR analyses we verified that even tRNAs possessing trancated secondary structures lacking either D arm or T arm could be folded into L-shape-like tertiary structures. 3) We succeeded to construct an in vitro translation system composed of only mt translation apparatuses. By using this system we proved that AUA isoleucine codon is translated as methionine by tRNA^<Met> having a modified base, f^6C at the anticodon first position. 4) Aminoacyl-tRNA synthetase, elongation factors, release factors, and formyltransferase were purified from bovine mitochondria, characterized, and their expression system was established in E.coli. We analyzed molecular mechanism of interaction between these factors and tRNA.

Report

(4 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • Research Products

    (52 results)

All Other

All Publications (52 results)

  • [Publications] N.Takeuchi: "Expression and Characterization of Bovine Mitochondrial Methiony1-tRNA Transformy1ase" J.Biochem.124. 1069-1071 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] I.Hayashi: "Higher-order Structure and Thermal Instability of Bobinc Mitochondrial tRNA ^<ser> UGA Investigated by Proton NMR Spectroscopy" J.Mol.Biol.284. 57-69 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Tomita: "7-Methylguanosine at the anticodom wobble position of squid mitochondrial tRNA ^<ser> GCU : molecular basis for assignment of AGA/AGG codons as serine in ivertebrate mitochondria" Biochimica et Biochysica Acta. 1399. 78-82 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Ohtsuki: "Stable Isotope-Edited NMR Analysis of Ascaris suum Mitochondrial tRNA ^<Met> Haring a TV-Replacement Loop" J.Biochem.124. 28-34 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] N.Takeuchi: "Mammalian Mitochondrial Methiony-tRNA Transformylase from Bovine Liver" Journal of Biological Chem.273-24. 15085-15090 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Kondow: "Assidian Mitochondorial tRNA ^<Met> Possessing Unique Structural Characteristics" Nucleosides & Nucleotides. 17(1-3). 531-539 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Matstuama: "A Novel Wobble Rule Found in Sarfish Mitochondria" Journal of Biological Chem.273-6. 3363-3368 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Y.Watanabe: "CDNA sequence of a translational elongation factor Ts homologue From caenorhabditis elegans : mitochondrial factor-spesific features found in the nematode homologue peptide" Biochimica et Biophysica Acta. 1353. 7-12 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] I.Hayashi: "Expression of bovine mitochondrial tRNA ^<Ser> GCU derivatives in Escherichia coli" Nucleic Scids Research. 25-17. 3503-3507 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] I.Hayashi: "Assignment of Imino Proton Signalw of G-C Base Paris and Magnesium Ion Binding : An NMR Study of Bovine Mitochondrial tRNA ^<Ser> GCU Lacking the Entire D Arm" J.Biochem.121. 1115-1122 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] N.Yamazaki: "Evolution of Pulmonate Gastropod Mitochondrial Genomes : Comparisons of Gene organizations of,Euhadra,Cepaea and Albinaria and Implications of Unusual tRNA Secondary Structures" Genetics. 145. 749-758 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Y.Watanabe: "Primary sequence of mitochondrial tRNA ^<Arg> of a nematode Ascaris suum : occurvence of adenosine at the first position of the anticodon" Biochimica et Biophysica Acta. 1350. 119-122 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Tomita: "Two nucleotides 5'-adjacent to the anticodon of rat cytoplasmic tRNA ^<Asp> are not edited" Biochimie. 78. 1001-1006 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Ohtsuki: "Automated Chemical Synthesis of Biologically Active tRNA Having a Sequence Correspondion to Ascaris suum Mitochondrial tRNA ^<Met> toward NMR Measurements" J.Biochem.120. 1070-1073 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] N.Nojima: "Pyridine-Promoted Factor^- and Energy-Free Petide sythesis Systems Prepared from Various Organisms Inoluding Prokaryote,Eukaryote,and Mitochondria" J.Biochem.119. 1076-1079 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Ohtsuki: "Preparation of biologically active Ascaris suum mitochondrial tRNA^<Met> with a TV-replacement loop by ligation of chemically synthesized RNA fragments." Nucleic Acids Research. 24-4. 662-667 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] N.Takeuchi: "Expression and Characterization of Bovine Mitochondrial Methionyl-tRNA Transformylase" J.Biochem.124. 1069-1071 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] I.Hayashi: "Higher-order Structure and Thermal Instability of Bobine Mitochondrial tRNA^<ser> UGA Investigated by Proton NMR Spectroscopy" J.Mol.Biol.284. 57-69 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Tomita: "7-Methylguanosine at the anticodon wobble position of squid mitochondrial tRNA^<ser> GCU : molecular basis for assignment of AGA/AGG codons as serine in ivertebrate mitochondria" Biochimica et Biophysica Acta. 1399. 78-82 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Ohtsuki: "Stable Iso tope-Edited NMR Analysis of Ascaris suum Mitochondrial tRNA^<Met> Having a TV-Replacement Loop" J.Biochem.124. 28-34 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] N.Takeuchi: "Mammalian Mitochondrial Methiony-tRNA Transformylase from Borine Liver" Journal of Biological Chem.273-24. 15085-15090 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Kondow: "Ascidian Mitochondrial tRNA^<Met> Possessing Unique Structural Characteristics" Nucleosides & Nucleotides. 17 (1-3). 531-539 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Matsuyama: "A Novel Wobble Rule Found in Starfish Mitochondria" Journal of Biological Chem.273-6. 3363-3368 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Y.Watanabe: "cDNA sequence of a translational clogation factor Ts homologue From Caenorhabditis elegans : mitochondrial factor-specific features found in the nematode homologue peptide" Biochimica et Biophysica Acta. 1353. 7-12 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] I.Hayashi: "Assignment of Imino Proton Signalw of G-C Base Pairs and Magnesium Ion Birding : An NMR Study of Bovine Mitochondrial tRNA^<ser> GCU Lacking the Entire D Arm" J.Biochem.121. 1115-1122 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] I.Hayashi: "Expression of bovine mitochondrial tRNA^<ser> GCU derivatives in Escherichia Coli" Nucleic Scids Research. 25-17. 3503-3507 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] N.Yamazaki: "Evolution of Pulmorate Gastropod Mitochondrial Genomes : Comparisons of Gene Organizations of Euhadra, Cepaea and Albinaria and Implications of Unusual tRNA Secondary Structures" Genetics. 145. 749-758 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Y.Watanabe: "Primary sequence of mitochondrial tRNA^<Arg> of a nematode Ascapis suum : occurrence of unmodified adenosime at the first position of the articodon" Biochimica et Biophysica Acta. 1350. 119-122 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] K.Tomita: "Two nucleotides s'-adjacent to the anticodon of rat cytoplasmic tRNA^<Arg> are not edited" Biochimie. 78. 1001-1006 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Ohtsuki: "Automated Chemical Synthesis of Biologically Active tRNA Having a Sequence Corresponding to Ascaris suum Mitochondrial tRNA^<Met> toward NMR Mesurements" J.Biochem.120. 1070-1073 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] N.Nojima: "Pyridine-Promoted Factor-and Energy-Free Petide Synthesis Systems Prepared from Various Organisms Including Prokaryote, Eukaryote, and Mitochondria" J.Biochem.119. 1076-1079 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Ohtsuki: "Preparation of biologically active Ascaris suum mitochondrial tRNA^<Met> with a TV-replacement loop by ligation of chemically synthesized RNA fragments" Nucleic Acids Research. 24-4. 662-667 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Hayashi, I., Kawai, G.& Watanabe, K.: "Higher-order structure and Thermal instability of bovine mitochondrial tRNA Series 4 investigated..." J.Mol.Biol.284. 57-69 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sasuga, J.et al.: "Gene Contents and Organization of a mitochondria DNA segments of the squid Loligo bleekeri" J.Mol.Evol.(印刷中).

    • Related Report
      1998 Annual Research Report
  • [Publications] Tomita, K, Ueda, T.& Watanabe, K.: "T-Methylquanosine at the anticodon wobble position of squid mitochondrial tRNA^<Ser>_<GCU>" Biochim.Biophys.Acta. 1339. 78-82 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ohtsuki et al.: "Stable isotope-edited NMR analysis of Ascaris suurm mitochondrial tRNA^<ret> having a TU-replace mart loop" J.Biochem.124. 28-34 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takeuchi, N.et al: "Mammalian mitochondrial methionyl-tRNA transformylase from bovine liver" J.Biol.Chem.273・24. 15085-15090 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kondow A.et al: "Ascidian mitochondrial tRNA^<ret> possessing unique structural characteristics" Nuclcosides & Nuclcotides. 17 (1-3). 531-539 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S.Matsuyama et al.: "A Novcl wobble rule found in starfish nifochondria" J.Biol.Chem.273・6. 3363-3368 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Watanabe et al.: "CDNA seguence of a transladinal eborgstion factor Ts humologue franc elegans : mitochondrial spacific feaornes" Biochim,Biodhys,Acfa. 1353. 7-12 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] I.Hayashi et al.: "Expression of bocine mitochondrial tRNA^<Ser> acu derivatives in Eschevi chia cali" Nucleic Acids Res. 25.17. 3503-3507 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] S.Yoshizawa et al: "GNA trinucleoride loop structures producing Extra-or bnarily stable DNA minihairpins" Biochemistry. 36・16. 4761-4767 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] I.Hayashi et al: "Asiignmend of Dmino profon signals of G-C base pais and magnesium ion binding" J.Biochem.121・6. 1115-1122 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Tomita et al: "Two nucleoti des S'-edjacent to the anticodon of rot cytoplasmic tRNA^<Asp> Cre not sdifed" Biochimie. 78. 1001-1006 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] 大澤著, 渡辺他訳: "違伝暗号の起渡と進化" 共立出版, 252 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Ohtsuki et al: "Automated Chemical synthesis of Biologically active tRNA having a sequence corresponding to Ascaris suum mitochondrial tRNA^<Met>" J.Biochem.120. 1070-1073 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Nojima et al: "Pyridine-psmotcd factor-and energy-free peptide synthesis systems prepared from Prokaryote eukaryote and mitochondria" J.Bioclem.119. 1076-1079 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] N.Yamazaki et al: "Evolution of Pulmonate Gastropod mitochondial genomes" Geneti cs. (印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Suzuki et al: "Polysemous codon-A codon coith multiple aminoacid assignmen caused by dual specifrcsty of tRNA identity" The EMBO J.(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Watanabe et al.: "An RNA fragment consisting of the P7 and P9.0 stems and the 3′-terminal guanosine of the tetralgmene group I intron" Nucl.Acids Res.24・7. 1337-1344 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Suzuki et al: "A new method for identifying the amino acid attached to a particular RNA in the cell" FEBS Lettems. 381. 195-198 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 三浦・渡辺編著: "バイオケミストリー" 昭晃堂, 221 (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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