1999 Fiscal Year Annual Research Report

RNA触媒機能の分子基盤

Research Project

Project/Area Number	09278103
Research Institution	Kyoto University
Principal Investigator	井上丹京都大学, 大学院・生命科学研究科, 教授 (40114855)
Co-Investigator(Kenkyū-buntansha)	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) 白石英秋京都大学, 大学院・生命科学研究科, 助教授 (90202118)
Keywords	RNA / リボザイム / グループIイントロン / 分子デザイン / ヘアピン型リボザイム / ハンマーヘッド型リボザイム / snRNP / G167
Research Abstract	・グループIイントロンRNAを材料としてリボザイムの反応の分子機構の解析を行い,触媒部位を含む最小の構造単位が,単純なpseudknot構造であることを明らかにした.また,分子デザインした人エタンパク質分子との複合体形成による同リボザイムの活性化に成功した.(井上) ・RNAの新規post synthetic modification法を確立した.また,それを用いてヘアピン型リボザイム内に,4-thio,2-thioおよび4-O-methyl uridine塩基を導入し,リボザイムの2つのドメインが相互作用するときに生ずるドメイン内の構造変化を明らかにした.(小松) ・ハンマーヘッド型リボザイムは生体内条件下では二価金属イオンを必須とする金属酵素であり,1個または2個の二価金属イオンが切断反応を触媒している.二価金属イオンが切断部位の非架橋酸素に結合して触媒作用を出していると一般的には信じられているが、その可能性を否定した。(多比良) ・U4/U6 snRNP構成因子である分裂酵母Prp1(pre-mRNA processing 1)と遺伝学的手法によって相互作用が検出されたPrp12をコードする遺伝子をクローニングした。その結果、splicing反応の初期において見られるcommitment complexからspliceosomeへの変換に必要なU4/U6snRNPとU2snRNP間の相互作用には、U2-U6snRNA間の水素結合による相互作用だけでなく、それらのsnRNAに結合している蛋白質間の相互作用も重要である可能性を示唆することに成功した。(谷) ・Volvox carteriの初期発生に関わる非コードRNAであるG167の解析を進めた.生殖細胞の分裂開始時に特徴的なこのRNAの発現の解析,対応する遺伝子のクローニング,また,他のVolvoxにおける同様の遺伝子の探索などを行った.(白石)

Research Products

(16 results)

All Other

All Publications (16 results)

[Publications] Tan Inoue: "Trans-activation of the Tetrahymena group I intron ribozyme via a non-native RNA-RNA interaction"Nucleic Acids Research. 27・7. 1650-1655 (1999)
[Publications] Tan Inoue: "A conserved motif in group IC3 introns is a new class of GNRA receptor"Nucleic Acids Research. 27・8. 1859-1865 (1999)
[Publications] Tan Inoue: "Characterization of P8 and J8/7 elements in the conserved core of the Tetrahymena group IO intron ribozyme"Biochemical and Biophysical Research Communications. 267. 85-90 (2000)
[Publications] Tan Inoue: "Requiremwnts for alternative forms of the activator domain, P5abc< in the Tetrahymena ribozyme"FEBS Letters. 462. 273-278 (2000)
[Publications] Komatsu, Y.: "Effects of a high-affinity antibody fragment on DNA polymerase reactions near a (6-4) photoproduct site"Photochem Photobiol. 69. 226-230 (1999)
[Publications] Komatsu, Y.: "Investigation of the recognition of an important uridine in an internal loop of a hairpin ribozyme prepared using post-synthetically modified oligonucleotides"Nucleic Acids Res.. 27. 4314-4323 (1999)
[Publications] Komatsu, Y.: "Conformational multiplicity of the antibody combining site of a monoclonal antibody specific for a (6-4) photoproduct"J. Mol. Biol.. 290. 731-740 (1999)
[Publications] Komatsu, Y.: "DNA binding mode of the Fab fragment of a monoclonal antibody specific for cyclobutane pyrimidine dimer"Nucleic Acids Res. 28. 944-951 (2000)
[Publications] Taira, K.: "tRNA Val-heterodimeric maxizymes with high potential as gene-inactivating agent : simultaneous cleavage at two sites in HIV-1 tat mRNA in cultured cells"Proc. Natl. Acad. Sci. USA. 96. 1886-1891 (1999)
[Publications] Taira, K.: "Extremely high and specific activity of DNA enzymes in cells with a Philadelphia chromosome"Chem. Biol.. 6. 237-250 (1999)
[Publications] Taira, K.: "CTAB-mediated enrichment for active forms of novel dimeric maxizymes"FEBS Lett.. 448. 67-74 (1999)
[Publications] Taira, K.: "Specificity of novel allosterically trans- and cis-activated connected maxiz ymes that are designed to suppress BCR-ABL expression"FEBS Lett.. 461. 77-85 (1999)
[Publications] Tokio Tani: "Characterization of the ptr6+ gene in fission yeast : A possible involvement of a transcriptional coactivator TAF in nucleocytoplasmic transport of mRNA"Genetics. 152. 869-880 (1999)
[Publications] Tokio Tani: "A "G" to "A" transition at the last nucleotide of exon 6 of the gc gene (868G-A) may result in either a splice or missensemutation in patients with X-linked severe combined immunodeficiency"Human Genet.. 104. 36-42 (1999)
[Publications] 谷時雄: "実験医学増刊「核-細胞質間輸送と小胞輸送」"mRNAの核外輸送機構. 52 (1999)
[Publications] 谷時雄: "蛋白質核酸酵素、増刊号「細胞核研究の最先端」"mRNAの核外輸送に関与する遺伝子群. 9 (1999)

1999 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・生命科学研究科, 教授 (40114855)

Research Products

[Publications] Tan Inoue: "Trans-activation of the Tetrahymena group I intron ribozyme via a non-native RNA-RNA interaction"Nucleic Acids Research. 27・7. 1650-1655 (1999)

[Publications] Tan Inoue: "A conserved motif in group IC3 introns is a new class of GNRA receptor"Nucleic Acids Research. 27・8. 1859-1865 (1999)

[Publications] Tan Inoue: "Characterization of P8 and J8/7 elements in the conserved core of the Tetrahymena group IO intron ribozyme"Biochemical and Biophysical Research Communications. 267. 85-90 (2000)

[Publications] Tan Inoue: "Requiremwnts for alternative forms of the activator domain, P5abc< in the Tetrahymena ribozyme"FEBS Letters. 462. 273-278 (2000)

[Publications] Komatsu, Y.: "Effects of a high-affinity antibody fragment on DNA polymerase reactions near a (6-4) photoproduct site"Photochem Photobiol. 69. 226-230 (1999)

[Publications] Komatsu, Y.: "Investigation of the recognition of an important uridine in an internal loop of a hairpin ribozyme prepared using post-synthetically modified oligonucleotides"Nucleic Acids Res.. 27. 4314-4323 (1999)

[Publications] Komatsu, Y.: "Conformational multiplicity of the antibody combining site of a monoclonal antibody specific for a (6-4) photoproduct"J. Mol. Biol.. 290. 731-740 (1999)

[Publications] Komatsu, Y.: "DNA binding mode of the Fab fragment of a monoclonal antibody specific for cyclobutane pyrimidine dimer"Nucleic Acids Res. 28. 944-951 (2000)

[Publications] Taira, K.: "tRNA Val-heterodimeric maxizymes with high potential as gene-inactivating agent : simultaneous cleavage at two sites in HIV-1 tat mRNA in cultured cells"Proc. Natl. Acad. Sci. USA. 96. 1886-1891 (1999)

[Publications] Taira, K.: "Extremely high and specific activity of DNA enzymes in cells with a Philadelphia chromosome"Chem. Biol.. 6. 237-250 (1999)

[Publications] Taira, K.: "CTAB-mediated enrichment for active forms of novel dimeric maxizymes"FEBS Lett.. 448. 67-74 (1999)

[Publications] Taira, K.: "Specificity of novel allosterically trans- and cis-activated connected maxiz ymes that are designed to suppress BCR-ABL expression"FEBS Lett.. 461. 77-85 (1999)

[Publications] Tokio Tani: "Characterization of the ptr6+ gene in fission yeast : A possible involvement of a transcriptional coactivator TAF in nucleocytoplasmic transport of mRNA"Genetics. 152. 869-880 (1999)

[Publications] Tokio Tani: "A "G" to "A" transition at the last nucleotide of exon 6 of the gc gene (868G-A) may result in either a splice or missensemutation in patients with X-linked severe combined immunodeficiency"Human Genet.. 104. 36-42 (1999)

[Publications] 谷時雄: "実験医学増刊「核-細胞質間輸送と小胞輸送」"mRNAの核外輸送機構. 52 (1999)

[Publications] 谷時雄: "蛋白質核酸酵素、増刊号「細胞核研究の最先端」"mRNAの核外輸送に関与する遺伝子群. 9 (1999)

井上丹京都大学, 大学院・生命科学研究科, 教授 (40114855)