2001 Fiscal Year Annual Research Report

転写装置の機能分化の全体像

Research Project

Project/Area Number	13480233
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)
Keywords	RNAポリメラーゼ / 転写調節 / 原核生物 / 真核生物 / 機能制御 / ゲノム / サブユニット / 蛋白-蛋白相互作用
Research Abstract	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と、我々は主張して来た。この理論を実証する目的で、原核生物、真核生物のモデル系を利用して研究を実施した。 1)大腸菌転写装置の機能分化:大腸菌シグマ因子7種類を純化して、RNAポリメラーゼコア酵素への結合の相対強度を決定した。また、特異抗体を作製し、免疫沈降法と二次元ゲル電気泳動法を併用して、各種培養条件で細胞内濃度を測定し、両者を合わせて、ホロ酵素濃度を理論的に予測することに成功した。一方、大腸菌で同定されている転写因子約150種類のうち、約50種類を発現純化し、それらの転写活性化または抑制機構を解明した。並行して、細胞内濃度の系統的測定を行った。 2)分裂酵母転写装置の機能分化:分裂酵母RNAポリメラーゼIIのサブユーツト全12種類の配置と集合機構を解析した。その上で、各サブユニットに直接接触する転写因子探索の系統的探索を開始した。普遍転写因子の幾つかや、サブユニット1のC端繰り返し構造を認識する蛋白リン酸化酵素と、脱リン酸化酵母の接点を見つけた。一方、各サブユニットを発現精製し、特異抗体を作製し、定量的ウエスタン法で、各サブユニツトの細胞内含量が測定できた。RNAポリメラーゼIIの細胞当りの数量は、やはりゲノム全遺伝子より少なかった。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を更に確信した。

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. Biocherm.. 268(3). 612-619 (2001)
[Publications] Mitobe, I: "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. Microbial.. 41(5). 1187-1198 (2001)
[Publications] Yasuno, K.: "Interaction of the C-terminal domain of the E. coli RNA polyrmerase 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 ofadenylate 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 (Fcpl)/TFIIF/RNA poly-merase II (Pol II) complex in Scizosaccharomyces pombe involves direct interaction between Fcpl 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"Mci. Microbial.. 43(2). 269-279 (2002)
[Publications] Colland, F.: "The interaction between δ^S, the stationary phase δ factor, and the core enzyme of Escherichia coli RNA polymertase"Genes Cells. (in press). (2002)
[Publications] Ishihama, A.: "Molecular recognition in gene transcription"Intenatl. 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 phosphoiylation 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

転写装置の機能分化の全体像

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. Biocherm.. 268(3). 612-619 (2001)

[Publications] Mitobe, I: "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. Microbial.. 41(5). 1187-1198 (2001)

[Publications] Yasuno, K.: "Interaction of the C-terminal domain of the E. coli RNA polyrmerase 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 ofadenylate 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 (Fcpl)/TFIIF/RNA poly-merase II (Pol II) complex in Scizosaccharomyces pombe involves direct interaction between Fcpl 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"Mci. Microbial.. 43(2). 269-279 (2002)

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

[Publications] Ishihama, A.: "Molecular recognition in gene transcription"Intenatl. 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 phosphoiylation 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)