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Mechanism of RNA function expression

Research Project

Project/Area Number 04272102
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Research InstitutionThe University of Tokyo

Principal Investigator

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

Co-Investigator(Kenkyū-buntansha) OHNO Mutsuhito  Kyoto University, Graduate School of Science, Research Associate, 大学院・理学研究科, 助手 (80201979)
NISHIKAWA Kazuya  Gifu University, Faculty of Engineering, Professor, 工学部, 教授 (60109262)
INOUE Tan  Kyoto University, Graduate School of Science, Professor, 大学院・理学研究科, 教授 (40114855)
SHIMURA Yoshiro  Biomolecular Engineering Research Institute, Director, 所長 (60025426)
YOKOYAMA Shigeyuki  University of Tokyo, Graduate School of Science, Professor, 大学院・理学系研究科, 教授 (00159229)
武藤 裕  東京大学, 理学部, 助手 (30192769)
坂本 博  神戸大学, 理学部, 助教授 (00187048)
Project Period (FY) 1992 – 1995
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥227,000,000 (Direct Cost: ¥227,000,000)
Fiscal Year 1995: ¥59,800,000 (Direct Cost: ¥59,800,000)
Fiscal Year 1994: ¥61,700,000 (Direct Cost: ¥61,700,000)
Fiscal Year 1993: ¥56,000,000 (Direct Cost: ¥56,000,000)
Fiscal Year 1992: ¥49,500,000 (Direct Cost: ¥49,500,000)
Keywordssplicing / alternative splicing / tRNA / modified nucleoside / base pairing / stable isotope labeling / NMR / sx1 protein / RNA / 機能構造 / nRNAスプライシング / リボザイム / ミトコンドリア / RNA結合蛋白質 / rRNA / mRNAスプライシング / ミトコンドリアtRNA
Research Abstract

This research group aims to elucidate the molecular mechanisms of RNA functions on the basis of their structures. It consists of two main groups, onestudying splicing mechanism and tRNA structure-functions, and another trying to establish methodology to enable structural analysis of RNAs by high-resolution NMR spactroscopy through labeling of RNAs site-specifically with stable isotopes. The splicing group has first found that U6sn RNA in spliceosomewhich is thought to be responsible for catalytic function, directly binds to mRNA precursor, the substrate for splicing reaction. The group also succeeded in elucidating regulation mechanism of the alternative splicing in Drosophila, which can be brought about by interactions between various gene products involved in it (such as tra and sxl proteins) and some specific sequences
in the substrate mRNA precursors. The tRNA group has concentrated on the structural analysis of animal mitochondrial (mt) tRNAs with unusual secondary structures and o … More n elucidating decoding mechanisms of tRNAs towards unusual genetic code. The group created a new idea that even unusual mt tRNAs can function on the ribosome as other usual tRNAs by a mechanism that the mutual distance and orientation between two functional sites of tRNA,the anticodon and the CCA end, can be preserved constant by special tertiary interactions within the mt tRNAs. The group also found that modified nucleosides in the anticodon first letter (or sometimes second letter) of mt tRNAs (such as 5-formylcytidine, 7-methylguanosine and pseudouridine) are directly involved in decoding of the unusual genetic code, by forming unusual codon-anticodon basepairings. The NMR group established the in virto synthesis method of RNA into the desired positions of which stable isotopes are introduced. By utilizing the method, the group succeeded in elucidating the recognition sites of guanine residue in groupI intron and the interaction of sx1 protein with a short RNA fragment which is recognized by the protein. The detailed structural analysis of the sx1 protein, itself was also carried out, which lead to elucidation of characteristic feature of amino acids residues involved in recognition of the RNA. Less

Report

(5 results)
  • 1996 Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (61 results)

All Other

All Publications (61 results)

  • [Publications] Watanabe,S.: "An RNA Fragment Consisting of the P7 and P9.0 Stems and the 3'-Terminal Guanosine of the Tetrahymena Group I Intron" Nucleic Acids Res.24. 1337-1344 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakamoto,K.: "NMR Studies of the Effects of the 5'-Phosphate Group on Conformational Properties of 5-Methylaminomethyluridine Found in the First Position of the Anticodon of Escherichia coli tRNA4Arg" Biochemistry. 35. 6533-6538 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Suzuki,T.: "A New Method for Identifying the Amino Acid Attached to a Particular RNA in the Cell" FEBS Letters. 381. 195-198 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ohtsuki,T.: "Preparation of Biologically Active Ascaris suum Mitochndrial tRNAMet with a TV-Replacement Loop by Ligation of Chemically Synthesized RNA Fragments" Nucleic Acids Res.24. 662-667 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] S.Watanabe, G.Kawai, Y.Muto, K.Watanabe, T.Inoue and S.Yokoyama: "An RNA Fragment Consisting of the P7 and P9.0 Stems and the 3'-Terminal Guanosine of the Tetrahymena Group I Intron." Nucleic Acids Res.24. 1337-1344 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Sakamoto, G.Kawai, S.Watanabe, T.Niimi, N.Hayashi, Y.Muto, K.Watanabe, T.Satoh, M.Sekine and S.Yokoyama: "NMR Studies of the Effects of the 5'-Phosphate Group on Conformational Properties of 5-Methylaminomethyluridine Found in the First Position of the Anticodon of Escherichia coli tRNA4Arg." Biochemistry. 35. 6533-6538 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Suzuki, T.Ueda and K.Watanebe: "A New Method for Identifying the Amino Acid Attached to a Particular RNA in the Cell." FEBS Letters. 381. 195-198 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Ohtsuki, G.Kawai, Y.Watanabe, K.Kita, K.Nishikawa and K.Watanabe: "Preparation of Biologically Active Ascaris suum Mitochondrial tRNAMet with a TV-Replacement Loop by Ligation of Chemically Synthesized RNA Fragments." Nucleic Acids Res.24. 662-667 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Asakawa,S.: "Nucleotide sequence and gene organization of the starfish Asterina pectinifera mitochondrial genome" Genetics. 140. 1047-1060 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nitta,I: "Template-dependent polypeptide synthesis in a factor-and energy-free translation system promoted by pyridine" J.Biochem. 118. 841-849 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nitta,I: "Factor-and energy-free peptide synthesis promoted by aromatic tertiary amines including nucleic acid-related compounds" J.Biochsm. 118. 850-854 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nitta,I: "A novelfactor -and energy-free translation system promoted by tertiary amines and its implications towards the origin of the traslation system" Viva Origino. 23. 209-222 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Takemoto,C.: "The ability of bovine mitochondrial transfer RNA(Met)to decode AUG and AUA codons." Biochimie. 77. 104-108 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Matsuo,M.: "Highly specific and efficient cleavage of squid tRNA^<Lys> catalyzed by magnesium ions." J.Biol.Chem. 270. 10097-10104 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Sekine,S.: "Major Identity Determinants in the Augmented D Helix of tRNA^<Glu> from Escherichia" J.Mol.Biol.256. 685-700 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nureki,O.: "Architectures of class-defining and specific domains of glutamyl-tRNAsynthetase" Science. 267. 1958-1965 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kigawa,T.: "Cell-Free Synthesis and Amino Acid-Selective Stable-Isotopic Labeling of Protein for NMR Analysis" J.Biomol.NMR. 6. 129-143 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Tateno,M.: "A three-dimensional strcuture model of the complex of glutamy1- tRNA synthetase and its cognate tRNA" FEBS lett.377. 77-81 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Seio,K.: "Synthesis and Properties of Conformationally Rigid Cyclouridylic Acid Having a Covalent Bonding linker between the Uracil 5-position and the 5′-Phoshate Group" Tetrahedron Letters. 36. 9515-9518 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Liu,J.: "The zinc-binding site of Escherichia coli glutamyl-tRNA synthetase is located in the scceptor-binding domain.Studies by extended x-ray absorption fine structure,molecular modeling,and site-directed mutagenesis." J.Biol.Chem. 270. 15162-15169 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Inoue,K: "Aspects of splice site selection in constitutive and alternative pre-mRNA splicing." Gene Expr.4. 177-182 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] J.Moriya: "A novel modified nucleoside found at the first position of the anticodon of methionine tRNA from bovine liver mitochondria." Biochemistry,. 33. 2234-2239 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Watanabe: "Unusual anticodon loop structure found in E.coli Lysine tRNA." Nucleic A cids Res.22. 79-87 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Suzuki: "Characterization of serine and leucine tRNAs in an asporogenic yeast Candida cylindracea and evolutionary implications of genes for tRNA^<Sor>CAG responsible for translation of a non-universal genetic code." Nucleic A cids Res.22. 115-123 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Wakita: "Higher-order structure of bovine mitochondrial tRNA^<Phe> lacking the “conserved" GG and TYCG sequences as inferred by enzymatic and chemical probing." Nucleic A cids Res.22. 347-353 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] I.Hirao: "Most compact hairpin-turn structure exerted by a short DNA fragment, d(GCGAAGC) in solution : an extraordinarily stable structure resistant to nucleases and heat." Nucleic A cids Res.22. 576-582 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] S.Yoshinari: "Existence of nuclear-encoded 5S-rRNA in bovine mitochondria." FEBS Lett.338. 137-142 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Takai: "Recognition of UUN Codons by Two Leucine tRNA Species from Escherichia coli" FEBS Lett.344. 31-34 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Hohsaka: "Site-Specific Incorporation of photofunctional nonnatural amino acids into a polypeptide through in vitro protein biosynthesis" FEBS Letters. 344. 171-174 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Sekine: "Chemical Synthesis and Properties of an Interresidually Cyclized Uridylyl(3'-5')uridine as a Model of tRNA U-Turn Structure Having a Sharp Bend" J. Am. Chem. Soc.116. 4469-4470 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Niimi: "Recognition of the Anticodon Loop of tRNA^<IIe>1 by Isoleucyl-tRNA Synthetase from Escherichia coli" Nucleotides and Nucleosides. 13. 1231-1237 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] H.Hosaka: "Sequence-specific Cleavage of Oligoribonucleotide Capable of Forming a Stem and Loop Structure" J. Biol. Chem.269. 20090-20094 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Tanaka: "Polypurine sequences within a downstream exon function as a splicing enhancer." Molecular & Cellular Biology. 14. 1347-1354 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] I.Nitta: "Template-Dependent Peptide Formation on Ribosomes Catalyzed by Pyridine." J. Biochem.115. 803-807 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] G.Kawai: "Conformational properties of a novel modified nucleoside, 5-formylcytidine, found at the first position of the anticodon of bovine mitochondrial tRNA^<Met>." Nucleosides and Nucleotides. 13. 1189-1199 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] S.Nakanishi: "A UGU Sequence in the Anticodon Loop Is a Minimum Requirement for Recognition by Escherichia coli tRNA-Guanine Transglycosylase" J. Biol. Chem.269. 32221-32225 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Y.Watanabe: "Higher-order structure of bovine mitochondrial tRNA^<Ser>UGA: chemical modificaiton and computer modeling" Nucleic A cids Res.22. 5378-5384 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] O.Nureki: "Molecular Recognition of the Identity-Determinant Set of Isoleucine Transfer RNA from Escherichia coli" J. Mol. Biol.236. 710-724 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] J.Moriya: "A Novel Modified Nucleoside Found at the First Position of the Anticodon of Methionine tRNA from Bovine Liver Mitochondria" Biochemistry. 33. 2234-2239 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K. P. Williams: "Selection of Novel Forms of a Functional Domain within the Tetrahymena Ribozyme" Nucleic A cids Res.22. 2003-2009 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K. P. Williams: "Two Universally Conserved Adenosines of the Group I Intron that are important for Selfsplicing but not for Core Catalytic Activity" J. Biochem.115. 126-130 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Ohama: "Non-universal decording of the leucine codon CUG in several Candida species" Nucleic Acids Res.21. 4039-4045 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] H.Asahara: "Recognition nucleotides of Escherichia coli tRNA^<Leu> and its elements facilitating discrimination from tRNA^<Ser> and tRNA^<Tyr>" J.Mol.Biol.231. 219-229 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Suzuki: "The gene for serine tRNA having anticodon sequence CAG in pathogenic yeast,Candida albicans" Nucleic Acids Res.21. 356 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] S.Yokobori: "Codon AGA and AGG are read as glycine in ascidian mitochondria" J.Mol.Evol.36. 1-8 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] O.Nureki: "Glutaminyl-tRNA synthetase from Thermus thermophilus HB8.Molecular cloning of the gltX gene and crystallization of the overproduced protein" Eur.J.Biochem.204. 465-472 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] A.Wakatabe: "The role of exon sequence in splice site selection" Genes & Development. 7. 407-418 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] H.Sakamoto: "Control of Drosophila Sex-lethal pre-mRNA splicing by its own femalespecific product" Nucleic Acids Res.20. 5533-5540 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Inoue: "Binding of the Drosophila transformer and transformer-2 proteins to the regulatory elements of doublesex primary transcript for sex-specific RNA processing" Proc.Natl.Acad.Sci.U.S.A.89. 8092-8096 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.P.Williams: "Two universally conserved adenosines of the group I intron that are important for self-splicing but not for core catalytic activity" Journal of Biochemistry,in press.(1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Ogawa: "Anticodon loop structure of Trulopsis utilis tRNA^<Val> and valine acceptance" J.Adv.Sci.5. 43-49 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Ohyama: "ATP and Mg^<2+> inos regulate both conformation and aminoacylation capacity of transfer RNA" J.Adv.Sci.5. 39-42 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Sakamoto: "A modified uridine in the first position of the anticodon of a minor species of arginie tRNA,the argU gene product,from Escherichia coli" Eur.J.Biochem.216. 369-375 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kumazawa,Y.: "Effect of the higher-order structure of tRNAs on the stability of hybrids with oligodeoxyribonucleotides:separation of tRNA by an efficient solution hybridization" Nucleic Acids Res.20. 2223-2232 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Ueda,T.: "The T-loop region of animal mitochondrial tRNA^<Ser>(AGY) is a main recognition site for homologous seryl-tRNA synthetase" Nucleic Acids Res.20. 2217-2222 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Osawa,S.: "Recent evidence for evolution of the genetic code" Microbiological Reviews. 56. 229-264 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Muramatsu,T.: "Recognition of the Nucleoside in the First Position of the Anticodon of Isoleucine tRNA by Isoleucyl-tRNA Synthetase from Escherichia coli" Nucleosides and Nucleotides. 11. 719-730 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Sawa,H.: "Association of U6 snRNA with the 5'-splice side region of pre-mRNA in the spliceosome" Genes & Development. 6. 244-254 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Yokogawa,T.: "Serine tRNA complementary to the nonuniversal serine codon CUG in Candida cylindracea:Evolutionary implications" Proc.Natl.Acad.Sci.USA. 89. 7408-7411 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kawai,G.: "Conformational rigidity of N^4-acetyl-2'-0-methylcytidine found in tRNA of extremely thermophilic archaebacteria(Archaea)" Nucleosides & Nucleotides. 11. 759-771 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kawai,G.: "Conformational rigidity of specific pryimidine residues in tRNA arises from posttranscriptional modifications that enhance steric interaction between the base and the 2'-hydroxyl group" Biochemistry. 31. 1040-1046 (1992)

    • Related Report
      1992 Annual Research Report

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

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