2000 年度実績報告書

転写装置の機能制御の分子基盤

研究課題

研究課題/領域番号	12147209
研究種目	特定領域研究(B)
研究機関	国立遺伝学研究所
研究代表者	石浜明国立遺伝学研究所, 分子遺伝研究系, 教授 (80019869)
研究分担者	藤田信之国立遺伝学研究所, 分子遺伝研究系, 助手 (90173434)
キーワード	生体金属 / 金属蛋白質 / 金属酵素 / 金属結合 / RNAポリメラーゼ / 転写装置 / 転写因子 / 転写制御
研究概要	RNAポリメラーゼは細菌では4種類のサブユニットからなり、亜鉛を含む多鎖金属酵素であり、また、活性中心にはマグネシウムが結合してRNA合成基質を捕捉する。RNAポリメラーゼは、多種多様な転写因子と相互作用をして転写対象の遺伝子の選択特性を変化させ、その結果、ゲノムの遺伝子の転写スペクトラムの変動がおきる。RNAポリメラーゼの特異性変換機構の解明を目的として、以下の研究を実施した。 (1)RNAポリメラーゼのサブユニット間相互作用、サブユニットと転写因子の相互作用のネットワーク、転写装置とDNA転写シグナルの相互作用を、鉄キレートFeBABEを蛋白に結合し、接触相手の蛋白の切断点を分析することで解明した。 (2)生物の金属結合蛋白質の全体像を理解する目的で、金属結合活性をもつ蛋白質の全てを同定する方法を開発した。本年度は、大腸菌全蛋白の中で、亜鉛結合活性をもつものを探索し、アミノ酸配列や質量分析から、多くの新規亜鉛結合蛋白質を同定した。

研究成果

(18件)

すべてその他

すべて文献書誌 (18件)

[文献書誌] Bown,J.A.: "Positioning of region 4 of the Escherichia coli RNA polymerase σ^<70> subunit by a transcription activator."J.Bact.. 182(10). 2982-2984 (2000)
[文献書誌] Fujita,N.: "Structural requirements for the interdomain linker of α Subunit of Escherichia coli RNA polymerase."Biochemistry. 39(20). 6243-6249 (2000)
[文献書誌] Hwang,J.-S.: "Expression of functional influenza virus RNA polymerase in the methylotrophic yeast Pichia pastoris."J.Virol.. 74(9). 4074-4084 (2000)
[文献書誌] Ishiguro,A.: "The Rpb6 subunit of fission yeast RNA polymerase II is a contact target of the transcription elongation factor TFII2."Mol.Cell.Biol.. 20(4). 1263-1270 (2000)
[文献書誌] Ishihama,A.: "Molecular anatomy of RNA polymerase using protein-conjugated metal probes with nuclease and protease activities."Chem.Comun.. 2000. 1091-1094 (2000)
[文献書誌] Katayama,A.: "Mapping of subunit-subunit contact surfaces on the β' subunit of Escherichia coli RNA polymerase."Jour.Biol.Chem.. 275(5). 3583-3592 (2000)
[文献書誌] Kimura.M.: "Involvement of multiple subunit-subunit contacts in the assembly of RNA polumerase II."Nucleic Acids.Res.. 28(4). 952-959 (2000)
[文献書誌] Maeda,H.: "Competition among seven Escherichiaa coli cs subunits : relative binding affinities to the core RNA polymerase."Nucleic Acids Res.. 28(18). 3497-3503 (2000)
[文献書誌] Maeda,H.: "Two extracytoplasmic function sigma subunits, σ^E and σ^<Fecl>, of Escherichia coli : Promoter selectivity and intracellular levels."J.Bacteriol.,. 182(4). 1181-1184 (2000)
[文献書誌] Ohnuma,M.: "A carboxy-terminal 16-amino-acid region of σ^<38> of Escherichia colt is important for transcription under high-salt conditions and sigma activities in vivo."J.Bact.. 182(16). 4628-4631 (2000)
[文献書誌] Otomo,T.: "Structural study of the N-terminal domain of the alpha subunit of Escherichia coli RNA polymerase solubilized with non-denaturing detergents."J.Biochem.. 128(2). 337-344 (2000)
[文献書誌] Ozoline,O.N.: "Transcription activation mediated by the carboxyl-terminal domain of the RNA polymerase s-subunit."J.Biol.Chem.. 275(2). 1119-1127 (2000)
[文献書誌] Shpakovski,G.V.: "Functional conservation of RNA polymerase II in fission and budding yeasts."J.Mol.BioL.. 295(5). 1119-1127 (2000)
[文献書誌] Talkuder,A.A.: "Two types of localization of the DNA-binding proteins within the Escherichia coli nucleoid."Genes to Cells. 5(8). 613-626 (2000)
[文献書誌] Wada,A.: "Growth phase-coupled changes of the ribosome profile in natural isolates and laboratory strains of Escherichia coli."J.Bact.. 182(19). 2893-2899 (2000)
[文献書誌] Yamamoto,K.: "Negative regulation or the bolAlp or Escherichia coli K- 12 by the transcription factor OmpR for osmolarity response genes."FEMS Microbiol.Lett.. 186(1). 257-262 (2000)
[文献書誌] Sakurai,H.: "Transcription organization and mRNA levels of the genes for all 12 subunits of the fission yeast RNA polymerase II."Genes to Cells. 6(1). 26-36 (2000)
[文献書誌] Ishihama,A.: "Annu.Rev.Microbiol."Functional modulation of Escherichia coli RNA polymerase.. 499-518 (2000)

2000 年度 実績報告書

転写装置の機能制御の分子基盤

研究代表者

石浜 明 国立遺伝学研究所, 分子遺伝研究系, 教授 (80019869)

研究成果

[文献書誌] Bown,J.A.: "Positioning of region 4 of the Escherichia coli RNA polymerase σ^<70> subunit by a transcription activator."J.Bact.. 182(10). 2982-2984 (2000)

[文献書誌] Fujita,N.: "Structural requirements for the interdomain linker of α Subunit of Escherichia coli RNA polymerase."Biochemistry. 39(20). 6243-6249 (2000)

[文献書誌] Hwang,J.-S.: "Expression of functional influenza virus RNA polymerase in the methylotrophic yeast Pichia pastoris."J.Virol.. 74(9). 4074-4084 (2000)

[文献書誌] Ishiguro,A.: "The Rpb6 subunit of fission yeast RNA polymerase II is a contact target of the transcription elongation factor TFII2."Mol.Cell.Biol.. 20(4). 1263-1270 (2000)

[文献書誌] Ishihama,A.: "Molecular anatomy of RNA polymerase using protein-conjugated metal probes with nuclease and protease activities."Chem.Comun.. 2000. 1091-1094 (2000)

[文献書誌] Katayama,A.: "Mapping of subunit-subunit contact surfaces on the β' subunit of Escherichia coli RNA polymerase."Jour.Biol.Chem.. 275(5). 3583-3592 (2000)

[文献書誌] Kimura.M.: "Involvement of multiple subunit-subunit contacts in the assembly of RNA polumerase II."Nucleic Acids.Res.. 28(4). 952-959 (2000)

[文献書誌] Maeda,H.: "Competition among seven Escherichiaa coli cs subunits : relative binding affinities to the core RNA polymerase."Nucleic Acids Res.. 28(18). 3497-3503 (2000)

[文献書誌] Maeda,H.: "Two extracytoplasmic function sigma subunits, σ^E and σ^<Fecl>, of Escherichia coli : Promoter selectivity and intracellular levels."J.Bacteriol.,. 182(4). 1181-1184 (2000)

[文献書誌] Ohnuma,M.: "A carboxy-terminal 16-amino-acid region of σ^<38> of Escherichia colt is important for transcription under high-salt conditions and sigma activities in vivo."J.Bact.. 182(16). 4628-4631 (2000)

[文献書誌] Otomo,T.: "Structural study of the N-terminal domain of the alpha subunit of Escherichia coli RNA polymerase solubilized with non-denaturing detergents."J.Biochem.. 128(2). 337-344 (2000)

[文献書誌] Ozoline,O.N.: "Transcription activation mediated by the carboxyl-terminal domain of the RNA polymerase s-subunit."J.Biol.Chem.. 275(2). 1119-1127 (2000)

[文献書誌] Shpakovski,G.V.: "Functional conservation of RNA polymerase II in fission and budding yeasts."J.Mol.BioL.. 295(5). 1119-1127 (2000)

[文献書誌] Talkuder,A.A.: "Two types of localization of the DNA-binding proteins within the Escherichia coli nucleoid."Genes to Cells. 5(8). 613-626 (2000)

[文献書誌] Wada,A.: "Growth phase-coupled changes of the ribosome profile in natural isolates and laboratory strains of Escherichia coli."J.Bact.. 182(19). 2893-2899 (2000)

[文献書誌] Yamamoto,K.: "Negative regulation or the bolAlp or Escherichia coli K- 12 by the transcription factor OmpR for osmolarity response genes."FEMS Microbiol.Lett.. 186(1). 257-262 (2000)

[文献書誌] Sakurai,H.: "Transcription organization and mRNA levels of the genes for all 12 subunits of the fission yeast RNA polymerase II."Genes to Cells. 6(1). 26-36 (2000)

[文献書誌] Ishihama,A.: "Annu.Rev.Microbiol."Functional modulation of Escherichia coli RNA polymerase.. 499-518 (2000)

2000 年度実績報告書

石浜明国立遺伝学研究所, 分子遺伝研究系, 教授 (80019869)