1997 年度実績報告書

RNA情報機能の分子基盤

研究課題

研究課題/領域番号	09278102
研究種目	重点領域研究
研究機関	東京大学
研究代表者	中村義一東京大学, 医科学研究所, 助教授 (40114590)
研究分担者	豊田春香東京大学, 医科学研究所, 講師 (10197973) 井上邦夫奈良先端科学技術院大学, 助手 (40252415) 松藤千弥東京慈恵会医科大学, 医学部, 助教授 (50192753) 口野嘉幸国立がんセター研究所, 部長(研究職) (60124418) 哀場弘二名古屋大学, 大学院・理学研究科, 教授 (20025662)
キーワード	RNA / 翻訳調節 / 転写後制御 / リボソーム / 分子擬態 / 終止コドン / mRNA動態 / RNA結合蛋白質
研究概要	平成9年度の研究成果は以下に整理し述べる。 1.翻訳調節におけるRNA情報機能の研究:(1)<リコーディング>動物細胞アンチザイムのフレームシフト信号の詳細な機能特性とポリアミンの促進作用点の同定、ならびに鳥類・魚類のアンチザイム遺伝子の分離によるフレームシフト信号の保存性を明らかにした。(2)<IRES>ポリオウィルスのIRES信号を組み込んだアデノウィルスベクター発現系を作成し、CATとLaczリポーターを利用して感染細胞レベルで強毒・弱毒ポリオウィルスのIRES活性を定量するシステムを作成した。(3)<翻訳終結>分裂酵母、テトラヒメナ、高度好熱菌等から解離因子をクローニングし、過剰発現、精製、遺伝解析系を作成した。(4)<発現調節>動物細胞のUAGサプレッサーグルタミンtRNA遺伝子の転写制御因子STGBPを同定し作用機作を明らかにした。 2.mRNAプロセシング機構の研究:(1)ゼブラフィッシュのElav/Hu型のRNA結合性因子(Etr-1,Etr-2)の同定と組織特異性、ならびにvasaホモログの分離とその生殖系列特異性を明らかにした。(2)Rho非依存的タ-ミネーターのステムループ構造がmRNAの安定化に普遍的に作用することを明らかにした。 3.tRNA情報機能の研究:(1)高度好熱菌のイソロイシルtRNA合成酵素について、L-イソロイシンおよびL-バリンとの複合体のX線結晶構造を2.5A分解能で明らかにし、校正機構を提示した。(2)解離因子によるtRNA分子擬態に係るアンチコドンならびにTステム擬態ドメインを体系的な変異分離作成により示唆した。大腸菌、サルモネラ菌、及び高度好熱菌の解離因子の結晶化を開始した。(3)カンジダ酵母で発見された多義語コドンtRNAが2種類のアミノ酸をチャージすることを証明した。

研究成果
(24件)

すべてその他

すべて文献書誌 (24件)

[文献書誌] Mita,K., Morimyo,M., Ito,K., Sugano,K., Ebihara,K., Hongo,E., Higashi,T., Hirayama,Y., Nakamura,Y.: "Comprehensive cloning of Schizosaccharamyces pombe genes encoding translation elougation factors." Gene. 187. 259-266 (1997)
[文献書誌] Ito,K., Nakamura,Y.: "Cloning and overexpression of polypeptide rerease factor 1 of Thermus thermophilus." Biochimie. 79. 287-292 (1997)
[文献書誌] Nakamura,Y., Wada,M.: "Molecular pathobiology and antigenic variation of Pneumocystis carinii." Adv.Parasitol. 41. 63-107 (1998)
[文献書誌] Wada,M., Nakamura,Y.: "Closing and overexpression of cell surface subtilisin-like proteases(SSP)of Pneumocystis carinii." J.Euk.microbiol. 54. 54S- (1997)
[文献書誌] Wada,M., Nakamura,Y.: "Hetero logous expression of Pneumocystis carinii genes in fission yeast." J.Euk.Microbiol.44. 55S-56S (1997)
[文献書誌] Moffat,J.G.Pel,II.J. Ito,K., Nakamura,Y., Tate,W.P.: "Escherichia coll release factor-3:resolving the paradox of a typical G protein atructure and atypical guanine nucleotide effects on function" RNA. 4. 47-54 (1998)
[文献書誌] Nakamura,Y.: "The major surface antigen of Pneumocystis carinii." FEMS Immun.Med.Microbiol.(in press). (1998)
[文献書誌] Nakamura,Y.: "Molecular microbiology of Pneumocystis carinii." Jpn J.Med.Mycol.(in press). (1998)
[文献書誌] Desgupta,S., Pernandez,L., Kameyama,L., Inada,T., Nakamura,Y., Pappas,A., Court,D.: "Genetic uncoupling of the dsRNA-binding and RNA-cleavage activities of the Escherichia coli endoribonuclease RNaseIII-the effect of dsRNA binding on gene expression." Mol.Microbiol.(in press). (1998)
[文献書誌] Ito,K., Ebihara,K., Nakamura,Y.: "Dissection of tRNA mimicry element of protein release factor eRF1 of fission yeast:eRF3 binding is not necessary for eRF1 function and cell viability." RNA. (in press). (1998)
[文献書誌] Kimata,K.: "CRP-cAMP plays a crucial role in glucose-lactose diauxie by activating the major glucose transporter gene in Escherichia coli." Proc.Natl.Acad.Sci.USA. 94. 12914-12919 (1997)
[文献書誌] Tagami,H.: "A common role of CRP in transcription activation:CRP acts transiently to stimulate events leading to open complex formation at a diverse set of promoters" EMBOJ.(in press). (1998)
[文献書誌] Hogema,B.: "Catabolite repression by glucose 6-phosphate,gluconate and lactose in Escherichia coli." Mol.Microbiol.24:857-867(1997). 24. 857-867 (1998)
[文献書誌] M.Nagane: "Application of antisense ribonucleic acid complementary to O6-methylguanine-deoxyribonucleic acid methyltansferase messenger nbonucleic acid for therapy of malignant gliomas" Neurosurgery. 41. 434-441 (1997)
[文献書誌] Matsufuji,S.: "Assignment of the human antizyme gene(OAZ)to chromosome 19p13.3 by fluorescence in situ hybridization" Genomics. 38. 102-104 (1996)
[文献書誌] Koguchi,K.: "Cloning and sequencing of a human cDNA encoding ornithine decarboxylase anti yme inhibites" Biochim.Biophys.Acta. 1353. 209-216 (1997)
[文献書誌] Hayashi.T.: "Characterization of the human antizyme gene." Gene. 203. 131-139 (1997)
[文献書誌] Tsuchihara,K.: "Specific Interaction of polypyrimidine tract-bingding protein with the extreme 3'-terminal structure of the Hepatitis C virus genome,the 3′X." J.Virol.71. 6720-6726 (1997)
[文献書誌] Artinger,M.: "Interaction of goosecoid and brachyury in Xenopus mesoderm patterning." Mech Dev. 65. 187-196 (1997)
[文献書誌] Kim,Y.: "NMR Analysis of the Hydrogen Bonding Interactions of the RNA-binding Domains of the Drosophila Sex-lethal Protein with Target RNA Fragments with Site-specific[3.15N]Uridine Substintion." Nucleic Acids Res.25. 1565-1569 (1997)
[文献書誌] S.Koshiba: "The Solution Structure of the Pleckstrin Homology Domain of Mouse Son-of-Sevenless 1 (mSos1)" J.Mol.Biol.269. 579-591 (1997)
[文献書誌] T.Nishinaka: "An Extended DNA Structure through Deoxyribose-base Stacking Induced by RecA Protein" Proc.Matl.Acad.Sci.USA. 94. 6623-6628 (1997)
[文献書誌] Y.Ito: "The ″Regional Polysterism″in the GTP-bound From of the Human c-Ha-Ras Protein." Biochemistry. 36. 9109-9119 (1997)
[文献書誌] M.Inoue: "A Characteristic Arrangement of Aromatie Amino Acid Residues in the Solution Structure of the Amino-terminal RNA-binding Domain of Drosophila Sex-lethal." J.Mol.Biol.272. 82-94 (1997)

1997 年度 実績報告書

RNA情報機能の分子基盤

研究代表者

中村 義一 東京大学, 医科学研究所, 助教授 (40114590)

研究成果

[文献書誌] Mita,K., Morimyo,M., Ito,K., Sugano,K., Ebihara,K., Hongo,E., Higashi,T., Hirayama,Y., Nakamura,Y.: "Comprehensive cloning of Schizosaccharamyces pombe genes encoding translation elougation factors." Gene. 187. 259-266 (1997)

[文献書誌] Ito,K., Nakamura,Y.: "Cloning and overexpression of polypeptide rerease factor 1 of Thermus thermophilus." Biochimie. 79. 287-292 (1997)

[文献書誌] Nakamura,Y., Wada,M.: "Molecular pathobiology and antigenic variation of Pneumocystis carinii." Adv.Parasitol. 41. 63-107 (1998)

[文献書誌] Wada,M., Nakamura,Y.: "Closing and overexpression of cell surface subtilisin-like proteases(SSP)of Pneumocystis carinii." J.Euk.microbiol. 54. 54S- (1997)

[文献書誌] Wada,M., Nakamura,Y.: "Hetero logous expression of Pneumocystis carinii genes in fission yeast." J.Euk.Microbiol.44. 55S-56S (1997)

[文献書誌] Moffat,J.G.Pel,II.J. Ito,K., Nakamura,Y., Tate,W.P.: "Escherichia coll release factor-3:resolving the paradox of a typical G protein atructure and atypical guanine nucleotide effects on function" RNA. 4. 47-54 (1998)

[文献書誌] Nakamura,Y.: "The major surface antigen of Pneumocystis carinii." FEMS Immun.Med.Microbiol.(in press). (1998)

[文献書誌] Nakamura,Y.: "Molecular microbiology of Pneumocystis carinii." Jpn J.Med.Mycol.(in press). (1998)

[文献書誌] Desgupta,S., Pernandez,L., Kameyama,L., Inada,T., Nakamura,Y., Pappas,A., Court,D.: "Genetic uncoupling of the dsRNA-binding and RNA-cleavage activities of the Escherichia coli endoribonuclease RNaseIII-the effect of dsRNA binding on gene expression." Mol.Microbiol.(in press). (1998)

[文献書誌] Ito,K., Ebihara,K., Nakamura,Y.: "Dissection of tRNA mimicry element of protein release factor eRF1 of fission yeast:eRF3 binding is not necessary for eRF1 function and cell viability." RNA. (in press). (1998)

[文献書誌] Kimata,K.: "CRP-cAMP plays a crucial role in glucose-lactose diauxie by activating the major glucose transporter gene in Escherichia coli." Proc.Natl.Acad.Sci.USA. 94. 12914-12919 (1997)

[文献書誌] Tagami,H.: "A common role of CRP in transcription activation:CRP acts transiently to stimulate events leading to open complex formation at a diverse set of promoters" EMBOJ.(in press). (1998)

[文献書誌] Hogema,B.: "Catabolite repression by glucose 6-phosphate,gluconate and lactose in Escherichia coli." Mol.Microbiol.24:857-867(1997). 24. 857-867 (1998)

[文献書誌] M.Nagane: "Application of antisense ribonucleic acid complementary to O6-methylguanine-deoxyribonucleic acid methyltansferase messenger nbonucleic acid for therapy of malignant gliomas" Neurosurgery. 41. 434-441 (1997)

[文献書誌] Matsufuji,S.: "Assignment of the human antizyme gene(OAZ)to chromosome 19p13.3 by fluorescence in situ hybridization" Genomics. 38. 102-104 (1996)

[文献書誌] Koguchi,K.: "Cloning and sequencing of a human cDNA encoding ornithine decarboxylase anti yme inhibites" Biochim.Biophys.Acta. 1353. 209-216 (1997)

[文献書誌] Hayashi.T.: "Characterization of the human antizyme gene." Gene. 203. 131-139 (1997)

[文献書誌] Tsuchihara,K.: "Specific Interaction of polypyrimidine tract-bingding protein with the extreme 3'-terminal structure of the Hepatitis C virus genome,the 3′X." J.Virol.71. 6720-6726 (1997)

[文献書誌] Artinger,M.: "Interaction of goosecoid and brachyury in Xenopus mesoderm patterning." Mech Dev. 65. 187-196 (1997)

[文献書誌] Kim,Y.: "NMR Analysis of the Hydrogen Bonding Interactions of the RNA-binding Domains of the Drosophila Sex-lethal Protein with Target RNA Fragments with Site-specific[3.15N]Uridine Substintion." Nucleic Acids Res.25. 1565-1569 (1997)

[文献書誌] S.Koshiba: "The Solution Structure of the Pleckstrin Homology Domain of Mouse Son-of-Sevenless 1 (mSos1)" J.Mol.Biol.269. 579-591 (1997)

[文献書誌] T.Nishinaka: "An Extended DNA Structure through Deoxyribose-base Stacking Induced by RecA Protein" Proc.Matl.Acad.Sci.USA. 94. 6623-6628 (1997)

[文献書誌] Y.Ito: "The ″Regional Polysterism″in the GTP-bound From of the Human c-Ha-Ras Protein." Biochemistry. 36. 9109-9119 (1997)

[文献書誌] M.Inoue: "A Characteristic Arrangement of Aromatie Amino Acid Residues in the Solution Structure of the Amino-terminal RNA-binding Domain of Drosophila Sex-lethal." J.Mol.Biol.272. 82-94 (1997)

1997 年度実績報告書

中村義一東京大学, 医科学研究所, 助教授 (40114590)