1995 年度実績報告書

RNAポリメラーゼの分子解剖:転写因子作用部位のマッピング

研究課題

研究課題/領域番号	06454672
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	転写調節 / RNAポリメラーゼ / 転写因子 / 分子集合 / 蛋白構造 / 分子間相互作用 / 分子解剖 / 遺伝解析
研究概要	遺伝子の転写制御の基本機構は、転写酵素RNAポリメラーゼが、さまざまの転写因子と相互作用した結果、転写をする対象遺伝子の選択を変換することによっていることが、明かになり初めている。転写因子とRNAポリメラーゼの分子間コミュニケーションの実体を解明する目的で、本研究では、RNAポリメラーゼ上の転写因子接点の同定を目指した。RNAポリメラーゼ各サブユニットに変異を導入することで、転写因子との接触不能となり、遺伝子制御が出来なくなることを指標にしたスクリーニングを行った。その結果、アルファサブユニットのC端領域に一群の転写因子(クラス1転写因子と命名)との接点を同定し、その領域各アミノ酸の役割を決定した。また、この領域を含む蛋白ドメインの三次元構造を決定し、転写因子と接触するアミノ酸の配置を決めた。その上に、この領域が、DNAのエンハンサーをも認識するサイトであることを実証した。これは、蛋白因子とDNA制御シグナルのいずれをも認識する、新しい蛋白機能域の発見となった。一方、RNAポイメラーゼの他のサブユニット、シグマ、ベタ-およびベタ-プライム上にも、他の転写因子との接点のクラスターが存在する予備的結果を得た。この間の研究で、大腸菌の約100種類の転写因子全ての接点を同定する方向の研究の基礎を築き、軌道に乗せた。

研究成果
(29件)

すべてその他

すべて文献書誌 (29件)

[文献書誌] Asano,Y.: "Molecular anatomy of influenza virus RNA polymerase:Photoaffinity labeling of PBI subunit with 8-azido GTP.21GC01:J.Biochem." 117. 677-682 (1995)
[文献書誌] Azuma,Y.: "Isolation of thermolabile RNA polymerase II from fission yeast Schizosaccharomyces pombe with mutations in the subunit 3 gene." J.Biochem.117. 216-220 (1995)
[文献書誌] Choy,H.: "Repression and activation of transcription by Gal and Lac repressors:Involvement of alpha-subunit of RNA polymerase." EMBOJ.14. 4523-4529 (1995)
[文献書誌] Ding,Q.: "Promoter selectivity of osmoregulated promoters between E^D and E^S holoenzymes." Mol.Microbiol.16. 649-656 (1995)
[文献書誌] Fukuchi,J.: "Decrease in cell viability due to the accumulation of spermidine in spermidine in spermidine acetyltransferease-deficient mutant of Escherichia coli." J.Biol.Chem.270. 18831-18835 (1995)
[文献書誌] Ishihama,A.: "Genetic strategies of bacteria for stationary-phase survival." Actinomycetologia. 9. 236-243 (1995)
[文献書誌] Kobayashi,M.: "Influenza virus PBI protein is the minimal and essential subunit of RNA polymerase." Arch.Virol.(1996)
[文献書誌] Kolb,A.: "Selectivity of the Escherichia coli RNA polymearase Eσ^<38> for overlapping promoters and ability to support CRP activation." Nucleic Acids Res.23. 819-826 (1995)
[文献書誌] Kumar,A.: "A partially functional 245 amino acid internal deletion derivative of Escherichia coli sigma-70." J.Bacteriol.177. 5193-5196 (1995)
[文献書誌] Kusano,S.: "Promoter selectivity of Escherichia coli RNA Polymerase Eσ^<70> and Eσ^<38> holoenzymes:Effect of DNA supercoiling." J.Biol.Chem.271. 1998-2004 (1996)
[文献書誌] Lawley,B.: "The TyrR protein of Escherichia coli is a class I transcription activator." J.Bacteriol.177. 238-241 (1995)
[文献書誌] Liu,X.: "The C-terminal region of the α subunit Escherichia coli RNA polymerase is required for transcription activation of the flagellar level II operons by the FlhD/FlhC complex." J.Bacteriol.177. 5186-5188 (1995)
[文献書誌] Jair,K.-W.: "ambidextrous transcription activation by SoxS:Requirement for the C-terminal domain of the RNA polymerase alpha subunit at a subset of supperoxide-inducible genes." Mol.Microbiol.19. 307-314 (1996)
[文献書誌] Jeon,Y.H.: "Solution structure of the C-terminal domain of RNA polymerase α subunit responsible for the contact with trans-acting transcription factors and cis-acting UP element of promoter." Science. 270. 1495-1997 (1995)
[文献書誌] Jishage,M.: "Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli:Intracellular levels of σ^<70> and σ^<38>." J.Bacteriol.177. 6832-6835 (1995)
[文献書誌] Kawagishi-Kobayashi,M.: "Mutational analysis of the RNase-like domain in subunit 2 of fission yeast RNA polymerase II." Mol.Gen.Genet.205. 1-5 (1996)
[文献書誌] Kimura,M.: "Functional map of the alpha subunit of Escherichia coli RNA polymerase:Insertion analysis of the amino-terminal assembly domain." J.Mol.Biol.248. 756-767 (1995)
[文献書誌] Kimura,M: "Functional map of the alpha subunit of Escherichia coli RNA polymerase:Amino acid substitutution within the amino-terminal assembly domain." J.Mol.Biol.254. 342-349 (1995)
[文献書誌] Nagai,S.: "The cyclic AMP receptor protein(CRP)gene function is required for expression of α (cell-bound)-type hemolysis in an avian pathogenic Escherichia coli strain." Infect.Immun.
[文献書誌] Nakagawa,Y.: "RNA polymerase PB2 subunit is not required for replication of the influenza virus genome,but is involved in capped RNA synthesis." J.Virol.69. 728-733 (1995)
[文献書誌] Negishi,T.: "Structural map of the alpha subunit of Escherichia coli RNA polymerase:Structural domains identified by proteolytic cleavage." J.Mol.Biol.248. 723-728 (1995)
[文献書誌] Shinkawa,H.: "Purification and characterization of RNA polymerase holoenzyme(Eσ^B)from vegetative-phase mycelia of Streptomyce griseus." J.Biochem.118. 488-495 (1995)
[文献書誌] Tanaka,K.: "Promoter determinants for Escherichia coli RNA polymerase holoenzyme containing σ^<38>(the rpoS gene product)." Nucleic Acids Res.23. 827-834 (1995)
[文献書誌] Tao,K.: "Mapping of the OxyR protein contact site in the C-terminal region of RNA polymerase α subunit." J.Bacteriol.177. 6740-6744 (1995)
[文献書誌] Toyoda,T.: "Molecular dissection of influenza virus RNA polymerase:PBI subunit alone is able to catalyze RNA synthesis.21GC25:Virus.Genes"
[文献書誌] Wada,A.: "Ribosome modulation factor:Stationary growth phase-specific inhibitor of ribosome functions from Escherichia coli." Biochem.Biophys.Res.Commun.214. 410-417 (1995)
[文献書誌] Yamagishi,M.: "Isolation of temperature-sensitive mutants of mRNA capping enzyme in Saccharomyces cerevisiae." Mol.Gen.Genet.249. 147-154 (1995)
[文献書誌] Jair,K.-W.: "Purification and regulatory properties of MarA protein:a tarnscriptional activator of Escherichia coli multiple antibiotic and superoxide resistance promoters." J.Bacteriol.177. 7100-7104 (1995)
[文献書誌] Fujita,N.: "Meth.Enzymol." (1996)

1995 年度 実績報告書

RNAポリメラーゼの分子解剖:転写因子作用部位のマッピング

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] Asano,Y.: "Molecular anatomy of influenza virus RNA polymerase:Photoaffinity labeling of PBI subunit with 8-azido GTP.21GC01:J.Biochem." 117. 677-682 (1995)

[文献書誌] Azuma,Y.: "Isolation of thermolabile RNA polymerase II from fission yeast Schizosaccharomyces pombe with mutations in the subunit 3 gene." J.Biochem.117. 216-220 (1995)

[文献書誌] Choy,H.: "Repression and activation of transcription by Gal and Lac repressors:Involvement of alpha-subunit of RNA polymerase." EMBOJ.14. 4523-4529 (1995)

[文献書誌] Ding,Q.: "Promoter selectivity of osmoregulated promoters between E^D and E^S holoenzymes." Mol.Microbiol.16. 649-656 (1995)

[文献書誌] Fukuchi,J.: "Decrease in cell viability due to the accumulation of spermidine in spermidine in spermidine acetyltransferease-deficient mutant of Escherichia coli." J.Biol.Chem.270. 18831-18835 (1995)

[文献書誌] Ishihama,A.: "Genetic strategies of bacteria for stationary-phase survival." Actinomycetologia. 9. 236-243 (1995)

[文献書誌] Kobayashi,M.: "Influenza virus PBI protein is the minimal and essential subunit of RNA polymerase." Arch.Virol.(1996)

[文献書誌] Kolb,A.: "Selectivity of the Escherichia coli RNA polymearase Eσ^<38> for overlapping promoters and ability to support CRP activation." Nucleic Acids Res.23. 819-826 (1995)

[文献書誌] Kumar,A.: "A partially functional 245 amino acid internal deletion derivative of Escherichia coli sigma-70." J.Bacteriol.177. 5193-5196 (1995)

[文献書誌] Kusano,S.: "Promoter selectivity of Escherichia coli RNA Polymerase Eσ^<70> and Eσ^<38> holoenzymes:Effect of DNA supercoiling." J.Biol.Chem.271. 1998-2004 (1996)

[文献書誌] Lawley,B.: "The TyrR protein of Escherichia coli is a class I transcription activator." J.Bacteriol.177. 238-241 (1995)

[文献書誌] Liu,X.: "The C-terminal region of the α subunit Escherichia coli RNA polymerase is required for transcription activation of the flagellar level II operons by the FlhD/FlhC complex." J.Bacteriol.177. 5186-5188 (1995)

[文献書誌] Jair,K.-W.: "ambidextrous transcription activation by SoxS:Requirement for the C-terminal domain of the RNA polymerase alpha subunit at a subset of supperoxide-inducible genes." Mol.Microbiol.19. 307-314 (1996)

[文献書誌] Jeon,Y.H.: "Solution structure of the C-terminal domain of RNA polymerase α subunit responsible for the contact with trans-acting transcription factors and cis-acting UP element of promoter." Science. 270. 1495-1997 (1995)

[文献書誌] Jishage,M.: "Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli:Intracellular levels of σ^<70> and σ^<38>." J.Bacteriol.177. 6832-6835 (1995)

[文献書誌] Kawagishi-Kobayashi,M.: "Mutational analysis of the RNase-like domain in subunit 2 of fission yeast RNA polymerase II." Mol.Gen.Genet.205. 1-5 (1996)

[文献書誌] Kimura,M.: "Functional map of the alpha subunit of Escherichia coli RNA polymerase:Insertion analysis of the amino-terminal assembly domain." J.Mol.Biol.248. 756-767 (1995)

[文献書誌] Kimura,M: "Functional map of the alpha subunit of Escherichia coli RNA polymerase:Amino acid substitutution within the amino-terminal assembly domain." J.Mol.Biol.254. 342-349 (1995)

[文献書誌] Nagai,S.: "The cyclic AMP receptor protein(CRP)gene function is required for expression of α (cell-bound)-type hemolysis in an avian pathogenic Escherichia coli strain." Infect.Immun.

[文献書誌] Nakagawa,Y.: "RNA polymerase PB2 subunit is not required for replication of the influenza virus genome,but is involved in capped RNA synthesis." J.Virol.69. 728-733 (1995)

[文献書誌] Negishi,T.: "Structural map of the alpha subunit of Escherichia coli RNA polymerase:Structural domains identified by proteolytic cleavage." J.Mol.Biol.248. 723-728 (1995)

[文献書誌] Shinkawa,H.: "Purification and characterization of RNA polymerase holoenzyme(Eσ^B)from vegetative-phase mycelia of Streptomyce griseus." J.Biochem.118. 488-495 (1995)

[文献書誌] Tanaka,K.: "Promoter determinants for Escherichia coli RNA polymerase holoenzyme containing σ^<38>(the rpoS gene product)." Nucleic Acids Res.23. 827-834 (1995)

[文献書誌] Tao,K.: "Mapping of the OxyR protein contact site in the C-terminal region of RNA polymerase α subunit." J.Bacteriol.177. 6740-6744 (1995)

[文献書誌] Toyoda,T.: "Molecular dissection of influenza virus RNA polymerase:PBI subunit alone is able to catalyze RNA synthesis.21GC25:Virus.Genes"

[文献書誌] Wada,A.: "Ribosome modulation factor:Stationary growth phase-specific inhibitor of ribosome functions from Escherichia coli." Biochem.Biophys.Res.Commun.214. 410-417 (1995)

[文献書誌] Yamagishi,M.: "Isolation of temperature-sensitive mutants of mRNA capping enzyme in Saccharomyces cerevisiae." Mol.Gen.Genet.249. 147-154 (1995)

[文献書誌] Jair,K.-W.: "Purification and regulatory properties of MarA protein:a tarnscriptional activator of Escherichia coli multiple antibiotic and superoxide resistance promoters." J.Bacteriol.177. 7100-7104 (1995)

[文献書誌] Fujita,N.: "Meth.Enzymol." (1996)

1995 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)