2001 Fiscal Year Annual Research Report

転写装置の機能制御の分子基盤

Research Project

Project/Area Number	12147209
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 / 転写調節 / 大腸菌 / 生体金属 / 機能制御 / 金属結合蛋白 / サブユニット / 蛋白-蛋白相互作用
Research Abstract	1)大腸菌RNAポリメラーゼは4種類のサブユニットからなり、亜鉛とマグネシウムを含む多鎖金属酵素である。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る。生物の遺伝情報転写包括制御は、転写装置の機能分化による、転写対象遺伝子選択スペクトラムの変化によっている。その分子機構の解明を目的として、大腸菌及び分裂酵母を利用して、サブユニット間、サブユニット-転写因子間の蛋白質相互作用ネットワークを系統的に解析した。予想されている転写開始シグマ因子7種類、転写因子約50種類について、精製し、転写装置の機能特異性への影響を調べた。原核生物、真核生物問わず、転写装置の遺伝子間の配分制御が、転写の包括制御の本質であることが示唆された。 2)生体内での転写装置の特異性変換を実証するモデル系として、大腸菌の環境中の金属への応答機構を解析した。 (1)亜鉛は、RNAポリメラーゼを含めて、蛋白-核酸相互作用、蛋白-蛋白相互作用のドメインに含まれていることが知らせている。大腸菌の亜鉛結合蛋白の全体像を知る目的で、二次元電気泳動による菌体全蛋白の分画と、放射性亜鉛結合活性を利用して、約50種類の新規亜鉛結合蛋白を同定した。金属結合活性を指標に各種金属結合蛋白が同定できる可能性がある。 (2)マグネシウムの不足時の環境からの取り込み、過剰時での環境への排出に関与する大腸菌遺伝子群を統括的に制御する転写因子PhoPを純化し、転写制御機構を明らかにした。また、Phop転写因子自体の、環境に相関した濃度変化を観測した。

Research Products
(24 results)

All Other

All Publications (24 results)

[Publications] Culham D.E: "The osmotic stress response and virulence in pyelonephritis isolates of Escherichia coli : contributions of RpoS, ProP, ProU and other systems"Microbiology. 147(6). 1657-1670 (2001)
[Publications] Gosh P.: "Escherichia coli RNA polymerase subunit ω and its N-terminal domain bind full length β' to facilitate incorporation into the α_2β subassemly"Eur. J. Biochem.. 268(7). 4621-4627 (2001)
[Publications] Honda A.: "Differential roles of viral RNA and cRNA in functional modulation of the influenza virus RNA polymerase"J. Biol. Chem.. 276(33). 31179-31185 (2001)
[Publications] Jishage M.: "Mapping of the Rsd contact site on the sigma 70 subunit of Escherichia coli RNA polymerase"J. Bacteriol.. 183(9). 2952-2956 (2001)
[Publications] Kimura M.: "Intracellular contents and assembly states of all 12 subunits of the RNA polymerase II in the fission yeast Schizosaccharomyces pombe"Eur. J. Biochem.. 268(3). 612-619 (2001)
[Publications] Mitobe J.: "Functional analysis of RNA polymerase II Rpb3 mutants of the fission yeast Schizosaccharomyces pombe"Curr. Genet.. 39(4). 210-221 (2001)
[Publications] Mitsuzawa H.: "Two WD repeat-containing TATA-binding protein-associated factors in fission yeast that suppress defects in the anaphase-promoting complex"J. Biol. Chem.. 276(29). 17117-17124 (2001)
[Publications] Ozoline O.N.: "Mode of DNA-protein interaction between the C-terminal domain of Escherichia coli RNA polymerase α subunit and T7D promoter UP element"Nucleic Acids Res.. 29(4). 4909-4919 (2001)
[Publications] Raj V.S.: "Properties of a revertant of Escherichia coli viable under spermidine accumulation : Increase in L-glycerol 3-phosphate"J. Bacteriol.. 183(15). 4493-4498 (2001)
[Publications] Sakurai H.: "Transcription organization and mRNA levels of the genes for all 12 subunits of the fission yeast RNA polymerase II"Genes Cells. 6(1). 26-36 (2001)
[Publications] Shin M.: "Repression of deoP2 in Escherichia coli by CytR : Conversion of a tanscription activator into a repressor"EMBO J.. 20(19). 5392-5399 (2001)
[Publications] Sujatha S.: "Functional complementation between mutations at two distant positions in Escherichia coli RNA polymerase as revealed by second-site suppression"Mol. Gen. Genet.. 264(5). 531-538 (2001)
[Publications] Wigneshweraraj S.R.: "In vitro roles of invariant helix-turm-helix motif residue R383 in σ^<54>(σ^N)"Nucleic Acids Res.. 29(7). 1163-1174 (2001)
[Publications] Wigneshweraraj S.R.: "Regulatory sequences in sigma 54 localise near the start of DNA melting"J. Mol. Biol.. 306(4). 681-701 (2001)
[Publications] Yamamoto K.: "Novel mode of transcription regulation by SdiA, an Eschreichia coli homologue of the quorum-sensing regulator"Mol. Microbiol.. 41(5). 1187-1198 (2001)
[Publications] Yasuno K.: "Interaction of the C-terminal domain of the E. coli RNA polymerase α subunit with the UP element : Recognizing the backbone structure in the minor groove surface"J. Mol. Biol.. 306(2). 213-225 (2001)
[Publications] Yoshida M.: "Polyamine enhancement of the synthesis of adenylate cyclase at the translational level and the consequential stimulation of the synthesis of the RNA polymerase σ^<28> subunit"J. Biol. Chem.. 276(19). 16289-16295 (2001)
[Publications] Kimura M.: "Formation of a carboxy-terminal domain-phosphatase(Fcp1)/TFIIF/RNA polymerase II(Pol II) complex in Scizosaccharomyces pombe involves direct interaction between Fcp1 and the Rpb4 subunit of Pol II"Mol. Cell. Biol.. 22(5). 1577-1578 (2002)
[Publications] Makinoshima H.: "Fractionation of Escherichia coli cell populations at different stages during growth transition to stationary phase"Mol. Microbiol.. 43(2). 269-279 (2002)
[Publications] Colland F.: "The interaction between σ^S, the stationary phase σ factor, and the core enzyme of Escherichia coli RNA polymertase"Genes Cells. (in press). (2002)
[Publications] Ishihama A.: "Molecular recognition in gene transcription"Internatl. J. Med. Biol. Frontiers. (in press). (2002)
[Publications] Sakurai H.: "Intracellular level of the RNA polymerase II in the fission yeast stays constant but the level of carboxyl-terminal domain phosphorylation varies depending on the phase and rate of cell growth"Genes Cells. (in press). (2002)
[Publications] Katayama A.: "Systematic search for zinc-binding proteins in Escherichia coli"Eur. J. Biochem.. (in press). (2002)
[Publications] Mitsuzawa H.: "Identification of histone H4-like TAF in Schizosaccharomyces pombe as a protein that interacts with WD repeart-containing TAF"Nucleic Acids Res.. (in press). (2002)

2001 Fiscal Year Annual Research Report

転写装置の機能制御の分子基盤

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] Culham D.E: "The osmotic stress response and virulence in pyelonephritis isolates of Escherichia coli : contributions of RpoS, ProP, ProU and other systems"Microbiology. 147(6). 1657-1670 (2001)

[Publications] Gosh P.: "Escherichia coli RNA polymerase subunit ω and its N-terminal domain bind full length β' to facilitate incorporation into the α_2β subassemly"Eur. J. Biochem.. 268(7). 4621-4627 (2001)

[Publications] Honda A.: "Differential roles of viral RNA and cRNA in functional modulation of the influenza virus RNA polymerase"J. Biol. Chem.. 276(33). 31179-31185 (2001)

[Publications] Jishage M.: "Mapping of the Rsd contact site on the sigma 70 subunit of Escherichia coli RNA polymerase"J. Bacteriol.. 183(9). 2952-2956 (2001)

[Publications] Kimura M.: "Intracellular contents and assembly states of all 12 subunits of the RNA polymerase II in the fission yeast Schizosaccharomyces pombe"Eur. J. Biochem.. 268(3). 612-619 (2001)

[Publications] Mitobe J.: "Functional analysis of RNA polymerase II Rpb3 mutants of the fission yeast Schizosaccharomyces pombe"Curr. Genet.. 39(4). 210-221 (2001)

[Publications] Mitsuzawa H.: "Two WD repeat-containing TATA-binding protein-associated factors in fission yeast that suppress defects in the anaphase-promoting complex"J. Biol. Chem.. 276(29). 17117-17124 (2001)

[Publications] Ozoline O.N.: "Mode of DNA-protein interaction between the C-terminal domain of Escherichia coli RNA polymerase α subunit and T7D promoter UP element"Nucleic Acids Res.. 29(4). 4909-4919 (2001)

[Publications] Raj V.S.: "Properties of a revertant of Escherichia coli viable under spermidine accumulation : Increase in L-glycerol 3-phosphate"J. Bacteriol.. 183(15). 4493-4498 (2001)

[Publications] Sakurai H.: "Transcription organization and mRNA levels of the genes for all 12 subunits of the fission yeast RNA polymerase II"Genes Cells. 6(1). 26-36 (2001)

[Publications] Shin M.: "Repression of deoP2 in Escherichia coli by CytR : Conversion of a tanscription activator into a repressor"EMBO J.. 20(19). 5392-5399 (2001)

[Publications] Sujatha S.: "Functional complementation between mutations at two distant positions in Escherichia coli RNA polymerase as revealed by second-site suppression"Mol. Gen. Genet.. 264(5). 531-538 (2001)

[Publications] Wigneshweraraj S.R.: "In vitro roles of invariant helix-turm-helix motif residue R383 in σ^<54>(σ^N)"Nucleic Acids Res.. 29(7). 1163-1174 (2001)

[Publications] Wigneshweraraj S.R.: "Regulatory sequences in sigma 54 localise near the start of DNA melting"J. Mol. Biol.. 306(4). 681-701 (2001)

[Publications] Yamamoto K.: "Novel mode of transcription regulation by SdiA, an Eschreichia coli homologue of the quorum-sensing regulator"Mol. Microbiol.. 41(5). 1187-1198 (2001)

[Publications] Yasuno K.: "Interaction of the C-terminal domain of the E. coli RNA polymerase α subunit with the UP element : Recognizing the backbone structure in the minor groove surface"J. Mol. Biol.. 306(2). 213-225 (2001)

[Publications] Yoshida M.: "Polyamine enhancement of the synthesis of adenylate cyclase at the translational level and the consequential stimulation of the synthesis of the RNA polymerase σ^<28> subunit"J. Biol. Chem.. 276(19). 16289-16295 (2001)

[Publications] Kimura M.: "Formation of a carboxy-terminal domain-phosphatase(Fcp1)/TFIIF/RNA polymerase II(Pol II) complex in Scizosaccharomyces pombe involves direct interaction between Fcp1 and the Rpb4 subunit of Pol II"Mol. Cell. Biol.. 22(5). 1577-1578 (2002)

[Publications] Makinoshima H.: "Fractionation of Escherichia coli cell populations at different stages during growth transition to stationary phase"Mol. Microbiol.. 43(2). 269-279 (2002)

[Publications] Colland F.: "The interaction between σ^S, the stationary phase σ factor, and the core enzyme of Escherichia coli RNA polymertase"Genes Cells. (in press). (2002)

[Publications] Ishihama A.: "Molecular recognition in gene transcription"Internatl. J. Med. Biol. Frontiers. (in press). (2002)

[Publications] Sakurai H.: "Intracellular level of the RNA polymerase II in the fission yeast stays constant but the level of carboxyl-terminal domain phosphorylation varies depending on the phase and rate of cell growth"Genes Cells. (in press). (2002)

[Publications] Katayama A.: "Systematic search for zinc-binding proteins in Escherichia coli"Eur. J. Biochem.. (in press). (2002)

[Publications] Mitsuzawa H.: "Identification of histone H4-like TAF in Schizosaccharomyces pombe as a protein that interacts with WD repeart-containing TAF"Nucleic Acids Res.. (in press). (2002)

石浜明国立遺伝学研究所, 分子遺伝研究系, 教授 (80019869)