1997 Fiscal Year Annual Research Report

RNA触媒機能の分子基盤

Research Project

Project/Area Number	09278103
Research Category	Grant-in-Aid for Scientific Research on Priority Areas
Research Institution	Kyoto University
Principal Investigator	井上丹京都大学, 大学院・理学研究科, 教授 (40114855)
Co-Investigator(Kenkyū-buntansha)	新田至東京大学, 工学系研究科, 助手 (30272404) 白石英秋京都大学, 大学院・理学研究科, 講師 (90202118) 谷時雄九州大学, 理学部, 助教授 (80197516) 多比良和誠筑波大学, 応用生物化学系, 教授 (10261778) 小松康雄北海道大学, 薬学部, 助手 (30271670)
Keywords	RNA / 酵素 / リボザイム / 分子設計 / 反応機構 / スプライシング / catalyticRNA / リボソーム
Research Abstract	・触媒活性をもつRNAの反応の分子機構の解析を,グループIイントロンRNAを材料として行い,イントロンRNAの機能構造単位の一つであるP2.1がtrans-activatorとして高効率でかつ新規なメカニズムで作用することを明らかにした.in vitro/in vivo selection法により新たにリボザイムを構築する計画を実施し、既知のRNAの高次構造を基盤としてリボザイムを作成するシステムの構築を試み,このシステムの作成に成功した.(井上) ・天然型のヘアピン型リボザイムより,3つのドメイン構造をもつ新規ヘアピン型リボザイムの構築に成功した.天然型よりも高い切断活性を示すヘアピン型リボザイムを構築した.(小松) ・ハンマーヘッド型リボザイムの反応機構として、一般的には"single-metal-ion"メカニズムが受け入れられているが、実際には"double-metal-ion"メカニズムであることを証明した。(多比良) ・分裂酵母mRNAスプライシング変異株の解析から、U4/U6snRNPの構成因子と考えられるPrp1が、G0/G1期変換に関与する因子と同一であることを明らかにした。(谷) ・大腸菌RNase Pは、活性中心をもつM1 RNAとC5タンパク質から成る複合体酵素である。C5タンパク質の機能を架橋実験によって解析し、これがM1 RNAとtRNAの位置関係を変化させる働きを持っていることを明らかにした。(白石) ・試験管内転写により調製された大腸菌由来の23SリボソームRNAを用いることにり、リボソームタンパク質を一切必要とすることなく、ペプチド結合生成反応を進行させることに成功し、リボソームRNA上に活性中心が存在することを証明した。(新田) ・新規のグアニン塩基結合RNAアプタマ-の単離に成功した.(坂本)

Research Products

(19 results)

All Other

All Publications (19 results)

[Publications] Ikawa,Y.: "Long-range interaction between the P2.1 and P9.1 peripheral domains of the Tetrahymena ribozyme." Nucleic Acids Research. 25. 1761-1765 (1997)
[Publications] Ikawa,Y.: "Idertification of the nucleotides in the A-rich bulge of the Tetrahymena ribozyme responsible for an efficient self-splicing." J.Biochem. 122. 878-882 (1997)
[Publications] Ikawa,Y.: "Trans-activation of the Tetrahymena ribozyme by its P2-2.1 domains" J.Biochem. 123. 528-533 (1998)
[Publications] Y.Naito: "P5abc of the Tetrahymena ribozyme consists of three functionally independent elements" RNA. (in press).
[Publications] Komatsu,Y.: "A new type of hairpin ribozyme consisting of three domains." Biochemistry. 36. 9935-9940 (1997)
[Publications] Komatsu,Y.: "Construction of hairpin ribozymes with a three-way junction." Bioorg.Med.Chem.5. 11063-11069 (1997)
[Publications] Urushiyama,S.: "The prp1+gene required for pre-mRNA splicing in Schizosaccharomyces pombe encodes a protein that contains TPR motifs and is similar to Prp6p of budding yeast." Genetics. 147. 101-115 (1997)
[Publications] J-H.Park: "Structure and expression of the gsa-1 gene encoding a G protein a (s) subunit in C. elegans" Gene. 194. 183-190 (1997)
[Publications] A.K.Azed: "Isolation and molecular characterization of mRNA transport mutants in Schizosaccharomyces pombe" Mol.Biol.Cell. 8. 825-841 (1997)
[Publications] Nitta,I.: "Possible involvement of Escherichia coli 23S ribosomal RNA in peptide bond formation." RNA. 4・3. 257-267 (1998)
[Publications] Nitta,I.: "A novel cell-free system for peptide synthesis driven by pyridine." Bio.Chem.(in press).
[Publications] D.Kiga,: "An RNA aptamer to the xanthine/guanine base with a distinctive mode of purine recognition" Nucleic Acids Research. (in press).
[Publications] Koizumi,M.: "Design and anti-HIV-1 activity of hammerhead and hairpin ribozymes containing a stable loop." Nucleosides & Nucleotides. (in press).
[Publications] Warashina,M.: "Entropically driven enhancement of cleavage activity of aDNA-armed hammerhead ribozyme : mechanism of action of hammerhead ribozymes." J.Org.Chem.62. 9138-9147 (1997)
[Publications] Fujita,S.: "Discrimination of a single base change in a ribozyme using the gene for dihydrofolate reductase as a selective marker in Escherichia coli." Proc.Natl.Acad.Sci.USA. 94. 391-396 (1997)
[Publications] Zhou,D.-M.: "Explanation by thedouble-metal-ion mechanism of catalysis for the differential metal ion effects on the cleavage rates of 5'-oxy and 5'-thio substrates by a hammerhead ribozyme." Proc.Natl.Acad.Sci.USA. 94. 14343-14348 (1997)
[Publications] Kuwabara,T.: "Comparison of the specificities and catalytic activities of hammerhead ribozymes and DNA enzymes with respect to the cleavage of BCR-ABL chimeric L6 (b2a2) mRNA." Nucleic Acids Res.25. 3074-3082 (1997)
[Publications] Imamura,O.: "Cloning and characterization of a human DEAH-box RNA helicase,a functional homolog of fission yeast Cdc28/Prp8." Nucleic Acids Res.(in press).
[Publications] 新田至, 上田卓也, 渡辺公綱: "リボソームとRNAワールド" 化学と生物 36(4), 92 (1998)

1997 Fiscal Year Annual Research Report

RNA触媒機能の分子基盤

Principal Investigator

井上 丹 京都大学, 大学院・理学研究科, 教授 (40114855)

Research Products

[Publications] Ikawa,Y.: "Long-range interaction between the P2.1 and P9.1 peripheral domains of the Tetrahymena ribozyme." Nucleic Acids Research. 25. 1761-1765 (1997)

[Publications] Ikawa,Y.: "Idertification of the nucleotides in the A-rich bulge of the Tetrahymena ribozyme responsible for an efficient self-splicing." J.Biochem. 122. 878-882 (1997)

[Publications] Ikawa,Y.: "Trans-activation of the Tetrahymena ribozyme by its P2-2.1 domains" J.Biochem. 123. 528-533 (1998)

[Publications] Y.Naito: "P5abc of the Tetrahymena ribozyme consists of three functionally independent elements" RNA. (in press).

[Publications] Komatsu,Y.: "A new type of hairpin ribozyme consisting of three domains." Biochemistry. 36. 9935-9940 (1997)

[Publications] Komatsu,Y.: "Construction of hairpin ribozymes with a three-way junction." Bioorg.Med.Chem.5. 11063-11069 (1997)

[Publications] Urushiyama,S.: "The prp1+gene required for pre-mRNA splicing in Schizosaccharomyces pombe encodes a protein that contains TPR motifs and is similar to Prp6p of budding yeast." Genetics. 147. 101-115 (1997)

[Publications] J-H.Park: "Structure and expression of the gsa-1 gene encoding a G protein a (s) subunit in C. elegans" Gene. 194. 183-190 (1997)

[Publications] A.K.Azed: "Isolation and molecular characterization of mRNA transport mutants in Schizosaccharomyces pombe" Mol.Biol.Cell. 8. 825-841 (1997)

[Publications] Nitta,I.: "Possible involvement of Escherichia coli 23S ribosomal RNA in peptide bond formation." RNA. 4・3. 257-267 (1998)

[Publications] Nitta,I.: "A novel cell-free system for peptide synthesis driven by pyridine." Bio.Chem.(in press).

[Publications] D.Kiga,: "An RNA aptamer to the xanthine/guanine base with a distinctive mode of purine recognition" Nucleic Acids Research. (in press).

[Publications] Koizumi,M.: "Design and anti-HIV-1 activity of hammerhead and hairpin ribozymes containing a stable loop." Nucleosides & Nucleotides. (in press).

[Publications] Warashina,M.: "Entropically driven enhancement of cleavage activity of aDNA-armed hammerhead ribozyme : mechanism of action of hammerhead ribozymes." J.Org.Chem.62. 9138-9147 (1997)

[Publications] Fujita,S.: "Discrimination of a single base change in a ribozyme using the gene for dihydrofolate reductase as a selective marker in Escherichia coli." Proc.Natl.Acad.Sci.USA. 94. 391-396 (1997)

[Publications] Zhou,D.-M.: "Explanation by thedouble-metal-ion mechanism of catalysis for the differential metal ion effects on the cleavage rates of 5'-oxy and 5'-thio substrates by a hammerhead ribozyme." Proc.Natl.Acad.Sci.USA. 94. 14343-14348 (1997)

[Publications] Kuwabara,T.: "Comparison of the specificities and catalytic activities of hammerhead ribozymes and DNA enzymes with respect to the cleavage of BCR-ABL chimeric L6 (b2a2) mRNA." Nucleic Acids Res.25. 3074-3082 (1997)

[Publications] Imamura,O.: "Cloning and characterization of a human DEAH-box RNA helicase,a functional homolog of fission yeast Cdc28/Prp8." Nucleic Acids Res.(in press).

[Publications] 新田至, 上田卓也, 渡辺公綱: "リボソームとRNAワールド" 化学と生物 36(4), 92 (1998)

井上丹京都大学, 大学院・理学研究科, 教授 (40114855)