1996 Fiscal Year Annual Research Report

RNAポリメラーゼの分子解剖:転写因子接点の同定と構造解析

Research Project

Project/Area Number	08458224
Research Category	Grant-in-Aid for Scientific Research (B)
Research Institution	National Institute of Genetics
Principal Investigator	石浜明国立遺伝学研究所, 分子遺伝研究系, 教授 (80019869)
Co-Investigator(Kenkyū-buntansha)	藤田信之国立遺伝学研究所, 分子遺伝研究系, 助手 (90173434)
Keywords	RNAポリメラーゼ / 転写因子 / DNA-蛋白質相互作用 / 蛋白質-蛋白質相互作用 / 分子集合 / 構造-機能相関 / 大腸菌 / 転写制御
Research Abstract	我々は、遺伝子の発現は、多種多様の転写調節因子がRNAポリメラーゼに作用し、その特異性(遺伝子認識選択特性)を変化する結果、転写対象の遺伝子が変化することによって制御されていることを明らかにしてきた。今回我々は、大腸菌RNAポリメラーゼを対象とし、大腸菌の存在すると予想される、約100種類の転写因子との接点を、4種のサブユニット上に同定し、さらに、転写因子との接触による、RNAポリメラーゼの機能特異性変換の機構を解明する目的で、本研究を実施した。研究は、先ず、大腸菌RNAポリメラーゼの変異体コレクションを作製し、世界各国の転写因子研究の専門家の強力を得て転写因子の供与をうけ、それぞれの転写因子の作用を受け付けなくなったRNAポリメラーゼ変異体を単離した。RNAポリメラーゼ上の変異の位置をDNA塩基配列から決定し、転写因子接点を推定した。遺伝解析から推定した接点を実証する目的で、さらに、RNAポリメラーゼと転写因子の接触を、直接に物理化学的、生化学的方法で解析した。約100種の転写因子の内、約30種については、接触するサブユニットを推定し、その内数種類については、アミノ酸配列の上で、接点を同定した。本研究によって、転写制御の分子機構の研究に、新たな突破口を切り開くことが出来た。

Research Products
(19 results)

All Other

All Publications (19 results)

[Publications] Artsimovitch,I.: "Transcription activation by the bacteriophage Mu Mor protein requires the C-terminal regions of both α and σ^<70> subunits of Escherichia coli RNA polymerase." J.Biol.Chem.271. 32343-32348 (1996)
[Publications] Belyaeva,T.: "Location of the C-terminal domain of the RNA polymerase alpha subunit in diggerent open complexes at the Escherichia coli galactose operon regulatory region." Nucleic Acids Res.24・12. 2243-2251 (1996)
[Publications] Eichenberger,P.: "The influence of the location of the activator binding site on the geometry of a transcriptional activation the geometry of a transcriptional activation complex in Eschericia coli." Biochemistry. 35・48. 15302-15312 (1996)
[Publications] Giladi,H.: "Identification of an UP element within the IHF binding site at the P_L1-P_L2 tandem promoter of bacteriophase λ." J.Mol.Biol.260. 484-491 (1996)
[Publications] Imamura,R.: "Identification of the cdpA gene encoding cyclic 3',5'-adenosine phosophate phosphodiesterase in Escherichia coli." J.Biol.Chem.271・41. 25423-25429 (1996)
[Publications] Jair,K.-W.: "Ambidextrous transcription activation by Soxs : Requirement for the C-terminal domain of the RNA polymerase alpha subunit at a subset of superoxide inducible genes." Mol.Microbiol.19・2. 307-314 (1996)
[Publications] Jair,K.-W.: "Transcription activation of promoters of the superoxide and multiple antibiotic resistance regulation by Rob,a binding protein of the Escherichia coli origin of chromosomal replication." J.Bacteriol.178・9. 2507-2513 (1996)
[Publications] Jishage.M.: "Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli : Intracellular levels of four species of sigma subunit under various growth conditons." J.Bacteriol.178・18. 5447-5451 (1996)
[Publications] Kimura,M.: "Subunit assembly in vivo of Escherichia coli RNA polymerase : Role of the amino-terminal assembly domain of alpha subunit." Genes to Cells. 1・6. 517-528 (1996)
[Publications] Kusano,S.: "Promoter selectivity of Escherichia coli RNA polymerase Eσ^<70> and Eσ^<38> holoenzymes : Effect of DNA supercoiling." J.Biol.Chem.271・4. 1998-2004 (1996)
[Publications] Luo,J.: "Molecular anatomy of the β' subunit of the E.coli RNA polymerase : Identification of region involved in polymerase assemblymerase : Identification of regions involved in polymerase assembly" Genes to Cells. 1・9. 819-828 (1996)
[Publications] Murakami,K.: "Transcription factor recognition surface on the RNA polymerase α subunit is involved in contact with the DNA enhancer element." EMBOJ.15・16. 4358-4367 (1996)
[Publications] Rajkumari,K.: "Effects of H-HS and potassium glutamate on σ^<S-> and σ^<70>-directed transcription in vitro from osmotically regulated P1 and P2 promoters of proU in Escherichia coli." J.Bacteriol.178・14. 4176-4181 (1996)
[Publications] Tang,Y.: "Upstream interactions at the lambda P_<RM> promoter are sequence-nonspecific and activate the promoter to a lesser extent than an introduced UP element of an rRna promoter." J.Bacteriol.178・23. 6945-6951 (1996)
[Publications] Greiner,D.P.: "Synthesis of the protein cutting reagent iron (S)-1-(p-bromoacetamidebenzym) ethylenediam inetetraacetate and conjucation to cysteine side chains." Bioconjugate Chemistry. 8・1. 44-48 (1997)
[Publications] Jishage,M.: "Variation in RNA polymerase sigma subunit composition within different stocks Escherichia coli strain W3110." J.Bacteriol.179・3. 959-963 (1997)
[Publications] Kusano,S.: "Promoter selectivity control of Escherichia coli RNA polyerase Eσ^<38> holoenzyme by trehalose." J.Bacteriol. (in press).
[Publications] Murakami,K.: "The two alpha subunits of Escherichia coli RNA polymerase are asymmetrically arranged and contact different halves of the DNA UP element." Proc.Nat.Acad.Sci.(in press).
[Publications] Negre,D.: "DNA flexibility of the UP element is a major determinant for transcriptional activation at the Escherichia coli acetate promoter." Nucleic Acids Res.25・4. 713-718 (1997)

1996 Fiscal Year Annual Research Report

RNAポリメラーゼの分子解剖:転写因子接点の同定と構造解析

Principal Investigator

石浜 明 国立遺伝学研究所, 分子遺伝研究系, 教授 (80019869)

Research Products

[Publications] Artsimovitch,I.: "Transcription activation by the bacteriophage Mu Mor protein requires the C-terminal regions of both α and σ^<70> subunits of Escherichia coli RNA polymerase." J.Biol.Chem.271. 32343-32348 (1996)

[Publications] Belyaeva,T.: "Location of the C-terminal domain of the RNA polymerase alpha subunit in diggerent open complexes at the Escherichia coli galactose operon regulatory region." Nucleic Acids Res.24・12. 2243-2251 (1996)

[Publications] Eichenberger,P.: "The influence of the location of the activator binding site on the geometry of a transcriptional activation the geometry of a transcriptional activation complex in Eschericia coli." Biochemistry. 35・48. 15302-15312 (1996)

[Publications] Giladi,H.: "Identification of an UP element within the IHF binding site at the P_L1-P_L2 tandem promoter of bacteriophase λ." J.Mol.Biol.260. 484-491 (1996)

[Publications] Imamura,R.: "Identification of the cdpA gene encoding cyclic 3',5'-adenosine phosophate phosphodiesterase in Escherichia coli." J.Biol.Chem.271・41. 25423-25429 (1996)

[Publications] Jair,K.-W.: "Ambidextrous transcription activation by Soxs : Requirement for the C-terminal domain of the RNA polymerase alpha subunit at a subset of superoxide inducible genes." Mol.Microbiol.19・2. 307-314 (1996)

[Publications] Jair,K.-W.: "Transcription activation of promoters of the superoxide and multiple antibiotic resistance regulation by Rob,a binding protein of the Escherichia coli origin of chromosomal replication." J.Bacteriol.178・9. 2507-2513 (1996)

[Publications] Jishage.M.: "Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli : Intracellular levels of four species of sigma subunit under various growth conditons." J.Bacteriol.178・18. 5447-5451 (1996)

[Publications] Kimura,M.: "Subunit assembly in vivo of Escherichia coli RNA polymerase : Role of the amino-terminal assembly domain of alpha subunit." Genes to Cells. 1・6. 517-528 (1996)

[Publications] Kusano,S.: "Promoter selectivity of Escherichia coli RNA polymerase Eσ^<70> and Eσ^<38> holoenzymes : Effect of DNA supercoiling." J.Biol.Chem.271・4. 1998-2004 (1996)

[Publications] Luo,J.: "Molecular anatomy of the β' subunit of the E.coli RNA polymerase : Identification of region involved in polymerase assemblymerase : Identification of regions involved in polymerase assembly" Genes to Cells. 1・9. 819-828 (1996)

[Publications] Murakami,K.: "Transcription factor recognition surface on the RNA polymerase α subunit is involved in contact with the DNA enhancer element." EMBOJ.15・16. 4358-4367 (1996)

[Publications] Rajkumari,K.: "Effects of H-HS and potassium glutamate on σ^<S-> and σ^<70>-directed transcription in vitro from osmotically regulated P1 and P2 promoters of proU in Escherichia coli." J.Bacteriol.178・14. 4176-4181 (1996)

[Publications] Tang,Y.: "Upstream interactions at the lambda P_<RM> promoter are sequence-nonspecific and activate the promoter to a lesser extent than an introduced UP element of an rRna promoter." J.Bacteriol.178・23. 6945-6951 (1996)

[Publications] Greiner,D.P.: "Synthesis of the protein cutting reagent iron (S)-1-(p-bromoacetamidebenzym) ethylenediam inetetraacetate and conjucation to cysteine side chains." Bioconjugate Chemistry. 8・1. 44-48 (1997)

[Publications] Jishage,M.: "Variation in RNA polymerase sigma subunit composition within different stocks Escherichia coli strain W3110." J.Bacteriol.179・3. 959-963 (1997)

[Publications] Kusano,S.: "Promoter selectivity control of Escherichia coli RNA polyerase Eσ^<38> holoenzyme by trehalose." J.Bacteriol. (in press).

[Publications] Murakami,K.: "The two alpha subunits of Escherichia coli RNA polymerase are asymmetrically arranged and contact different halves of the DNA UP element." Proc.Nat.Acad.Sci.(in press).

[Publications] Negre,D.: "DNA flexibility of the UP element is a major determinant for transcriptional activation at the Escherichia coli acetate promoter." Nucleic Acids Res.25・4. 713-718 (1997)

石浜明国立遺伝学研究所, 分子遺伝研究系, 教授 (80019869)