1994 Fiscal Year Annual Research Report

RNA機能の発現機構

Research Project

Project/Area Number	04272102
Research Institution	The University of Tokyo
Principal Investigator	渡辺公綱東京大学, 工学部, 教授 (00134502)
Co-Investigator(Kenkyū-buntansha)	大野睦人京都大学, 理学部, 助手 (80201979) 西川一八東京工業大学, 生命理工学部, 助教授 (60109262) 井上丹京都大学, 理学部, 教授 (40114855) 志村令郎京都大学, 理学部, 教授 (60025426) 横山茂之東京大学, 大学院・理学系研究科, 教授 (00159229)
Keywords	RNA / 機能構造 / mRNAスプライシング / リボザイム / tRNA / ミトコンドリア / RNA結合蛋白質 / NMR
Research Abstract	スプライシンググループ (1)mRNAスプライシングの分子機構ポリプリン配列がスプライシング反応のエンハンサーとして機能することを明らかにした。また、スプライシング反応に関与するRNAヘリカーゼをコードするものと考えられるcDNA配列をHeLa細胞から分離同定した。さらに、80KDaの核由来キャップ構造結合タンパク質を分離し、このタンパク質をコードするcDNAの全塩基配列を決定した。 (2)リボザイムの機能構造の研究フットプリティング法、反応速度論による解析などを通じて、二分子のRNAから構成されるリボザイムにおけるRNA間の相互作用を解明した。一方、in vitro selection法によって異なるメカニズムでリボザイムの活性化をおこなうRNAを得た。 tRNAグループ (1)異常構造をとるミトコンドリアtRNAの機能構造Dアームを殆ど欠失した牛ミトコンドリアのtRNA^<Ser>_<GCU>についてはT7RNAポリメラーゼを用いたin vitroの転写系によって、一方、Tアームを欠失した線虫ミトコンドリアのメチオニンtRNAについては化学合成および酵素的連結によって、それぞれ数mgの試料を調製し、NMRスペクトルを測定した。さらに、それらの安定同位体標識および位置選択的なイノシン残基の導入も行った。 (2)分子整形術によるtRNAの位置選択的安定同位体標識分子整形術によって、変則暗号を解読するCandida cylindracea のtRNA^<Ser>_<CAG>の33および34位に安定同位体標識ヌクレオチドを導入し、NMRスペクトルを測定した。 NMRグループ (1)RNAの安定同位体標識 RNAを安定同位体標識する方法をほぼ確立した。 (2)RNAの機能構造多様なRNA(snRNA U6,グアニン・キサンチン結合RNA,group I intronモデルRNA,28SrRNA GTPaseドメイン)について、NMRなどによる解析を行った。 (3)RNAとRNA結合タンパク質の相互作用 Sx1タンパク質の2つのRNA結合ドメイン(d1,d2)がともに特異的なRNA結合能を持つことを見いだした。さらに、グルタミルtRNA合成酵素によるグルタミン酸tRNAの認識機構を明らかにした。

Research Products

(20 results)

All Other

All Publications (20 results)

[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)
[Publications] K.Watanabe: "Unusual anticodon loop structure found in E.coli Lysine tRNA." Nucleic A cids Res.22. 79-87 (1994)
[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)
[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)
[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)
[Publications] S.Yoshinari: "Existence of nuclear-encoded 5S-rRNA in bovine mitochondria." FEBS Lett.338. 137-142 (1994)
[Publications] K.Takai: "Recognition of UUN Codons by Two Leucine tRNA Species from Escherichia coli" FEBS Lett.344. 31-34 (1994)
[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)
[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)
[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)
[Publications] H.Hosaka: "Sequence-specific Cleavage of Oligoribonucleotide Capable of Forming a Stem and Loop Structure" J. Biol. Chem.269. 20090-20094 (1994)
[Publications] K.Tanaka: "Polypurine sequences within a downstream exon function as a splicing enhancer." Molecular & Cellular Biology. 14. 1347-1354 (1994)
[Publications] I.Nitta: "Template-Dependent Peptide Formation on Ribosomes Catalyzed by Pyridine." J. Biochem.115. 803-807 (1994)
[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)
[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)
[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)
[Publications] O.Nureki: "Molecular Recognition of the Identity-Determinant Set of Isoleucine Transfer RNA from Escherichia coli" J. Mol. Biol.236. 710-724 (1994)
[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)
[Publications] K. P. Williams: "Selection of Novel Forms of a Functional Domain within the Tetrahymena Ribozyme" Nucleic A cids Res.22. 2003-2009 (1994)
[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)

1994 Fiscal Year Annual Research Report

RNA機能の発現機構

Principal Investigator

渡辺 公綱 東京大学, 工学部, 教授 (00134502)

Research Products

[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)

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

[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)

[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)

[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)

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

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

[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)

[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)

[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)

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

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

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

[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)

[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)

[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)

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

[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)

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

[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)

渡辺公綱東京大学, 工学部, 教授 (00134502)