1997 Fiscal Year Annual Research Report

RNA情報機能の分子基盤

Research Project

Project/Area Number	09278102
Research Category	Grant-in-Aid for Scientific Research on Priority Areas
Research Institution	The University of Tokyo
Principal Investigator	中村義一東京大学, 医科学研究所, 助教授 (40114590)
Co-Investigator(Kenkyū-buntansha)	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)
Keywords	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
Research Abstract	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。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] 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)
[Publications] Ito,K., Nakamura,Y.: "Cloning and overexpression of polypeptide rerease factor 1 of Thermus thermophilus." Biochimie. 79. 287-292 (1997)
[Publications] Nakamura,Y., Wada,M.: "Molecular pathobiology and antigenic variation of Pneumocystis carinii." Adv.Parasitol. 41. 63-107 (1998)
[Publications] Wada,M., Nakamura,Y.: "Closing and overexpression of cell surface subtilisin-like proteases(SSP)of Pneumocystis carinii." J.Euk.microbiol. 54. 54S- (1997)
[Publications] Wada,M., Nakamura,Y.: "Hetero logous expression of Pneumocystis carinii genes in fission yeast." J.Euk.Microbiol.44. 55S-56S (1997)
[Publications] 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)
[Publications] Nakamura,Y.: "The major surface antigen of Pneumocystis carinii." FEMS Immun.Med.Microbiol.(in press). (1998)
[Publications] Nakamura,Y.: "Molecular microbiology of Pneumocystis carinii." Jpn J.Med.Mycol.(in press). (1998)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Hogema,B.: "Catabolite repression by glucose 6-phosphate,gluconate and lactose in Escherichia coli." Mol.Microbiol.24:857-867(1997). 24. 857-867 (1998)
[Publications] 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)
[Publications] Matsufuji,S.: "Assignment of the human antizyme gene(OAZ)to chromosome 19p13.3 by fluorescence in situ hybridization" Genomics. 38. 102-104 (1996)
[Publications] Koguchi,K.: "Cloning and sequencing of a human cDNA encoding ornithine decarboxylase anti yme inhibites" Biochim.Biophys.Acta. 1353. 209-216 (1997)
[Publications] Hayashi.T.: "Characterization of the human antizyme gene." Gene. 203. 131-139 (1997)
[Publications] 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)
[Publications] Artinger,M.: "Interaction of goosecoid and brachyury in Xenopus mesoderm patterning." Mech Dev. 65. 187-196 (1997)
[Publications] 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)
[Publications] S.Koshiba: "The Solution Structure of the Pleckstrin Homology Domain of Mouse Son-of-Sevenless 1 (mSos1)" J.Mol.Biol.269. 579-591 (1997)
[Publications] T.Nishinaka: "An Extended DNA Structure through Deoxyribose-base Stacking Induced by RecA Protein" Proc.Matl.Acad.Sci.USA. 94. 6623-6628 (1997)
[Publications] Y.Ito: "The ″Regional Polysterism″in the GTP-bound From of the Human c-Ha-Ras Protein." Biochemistry. 36. 9109-9119 (1997)
[Publications] 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 Fiscal Year Annual Research Report

RNA情報機能の分子基盤

Principal Investigator

中村 義一 東京大学, 医科学研究所, 助教授 (40114590)

Research Products

[Publications] 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)

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

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

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

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

[Publications] 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)

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

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

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

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

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

[Publications] 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)

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

[Publications] 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)

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

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

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

[Publications] 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)

中村義一東京大学, 医科学研究所, 助教授 (40114590)