2003 Fiscal Year Annual Research Report

大腸菌転写制御ネットワークの包括的解析

Research Project

Project/Area Number	15013221
Research Institution	Nagoya University
Principal Investigator	饗場浩文名古屋大学, 大学院・生命農学研究科, 助教授 (60211687)
Co-Investigator(Kenkyū-buntansha)	山本兼由近畿大学, 農学部, 助手 (40351580) 大島拓奈良先端科学技術大学院大学, 遺伝子教育センター, 助手 (50346318) 饗庭弘二名古屋大学, 大学院・理学研究科, 教授 (20025662)
Keywords	2成分制御系 / 大腸菌 / 転写制御 / マイクロアレイ / レスポンスレギュレーター / ヒスチジンキナーゼ / ストレス応答 / 情報伝達
Research Abstract	大腸菌レスポンスレギュレーター高発現株における遺伝子発現の網羅的解析 2成分制御系のターゲット遺伝子を検索する目的で、全レスポンスレギュレーターの高発現株を作製し、そのマイクロアレイ解析をおこなった。ここで得られたデータを、すでに取得した2成分制御系の欠失株でのマイクロアレイデータと総合することにより、レスポンスレギュレーターにより活性化される遺伝子群、抑制される遺伝子群の候補を抽出した。その結果、機能未知の2成分制御系であるYpdABは糖の取り込みと利用に関わる遺伝子群を、BasSRは薬剤排出ポンプを制御していること示された。その他の候補遺伝子群についても順次、遺伝学的、生化学的解析を進行中である。また、すでに夾膜多糖の合成制御に関わることが示されていたRcsC-YojN-RcsBに関しては、新たにストレスシグナルを同定した(グルコース、亜鉛、低温)。加えて、RcsC-YojN-RcsBの上位には、2成分制御系PhoPQが存在し、前述の亜鉛シグナルは、PhoPQ系を介してRcsC-YojN-RcsB系に伝達されることも見出した。これは、2成分制御系間ネットワーク(カスケード)の1例と考えることができる。 2成分制御系間のクロストークを網羅的に解析るために、全てのヒスチジンキナーゼならびにレスポンスレギュレーターを精製し、in vitroにおいてリン酸化反応を行った。その結果、9種類のヒスチジンキナーゼは本来のペアー以外のレスポンスレギュレーターもリン酸化することを見出した。こらら候補については、in vitroにおいても生理学的に意味のあるクロストークしている可能性が高いので、解析をさらに進めている。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Watanabe, T.: "Isolation and characterization of inhibitors of the essential histidine kinase, YycG in Bacillus subtilis and Staphylococcus aureus."J.Antibiotics. 56. 1045-1052 (2003)
[Publications] Eguchi, Y.: "Transcriptional regulation of drug efflux genes by EvgAS, a two-component system in Escherichia coli."Microbiology. 149. 2819-2828 (2003)
[Publications] Aketani, S.: "Role of a Nonnatural b-C-Nucleotide Unit in DNA as Template for DNA and RNA Synthesis and as Substrate for Nucleolytic Digestion."Eur.J.Phram.Sci.. 20. 43-51 (2003)
[Publications] Minagawa, S.: "Identification and molecular characterization of the Mg2+ stimulon of Escherichia coli."J.Bacteriol.. 185. 3696-3702 (2003)
[Publications] Gowrishankar, J.: "In Vitro Properties of RpoS (sigma(S)) Mutants of Escherichia coli with Postulated N-Terminal Subregion 1.1 or C-Terminal Region 4 Deleted."J.Bacteriol.. 185. 2673-2679 (2003)
[Publications] Yamamoto, K.: "Two different modes of transcription repression of the Escherichia coli acetate operon by Ic1R"Mol.Microbiol.. 47. 183-194 (2003)
[Publications] 中道範人: "分裂酵母における胞子形成制御するHis-Aspリン酸リレー情報伝達"日本農芸化学会誌. 78. 50-51 (2004)
[Publications] Yamamoto, Y.: "SsrA-mediated trans-translation plays a role in mRNA quality control by facilitating degradation of truncated mRNAs."RNA. 9. 408-418 (2003)
[Publications] EI-Kazzaz, W.: "Metabolic block at early stages of glycolytic pathway activates the Rcs phosphorelay system via increased synthesis of dTDP-glucose in Escherichia coli."Mol.Microbiol.. 51. 1117-1128 (2004)
[Publications] Sunohara, T.: "Nascent peptide-mediated ribosome stalling at a stop codon induces mRNA cleavage resulting in nonstop mRNA that is recognized by tmRNA."RNA. 10. 378-386 (2004)
[Publications] Eguchi, Y.: "A signal transduction cascade between EvgA/EvgS and PhoP/PhoQ two-component systems of Escherichia coli."J.Bacteriol.. (印刷中). (2004)
[Publications] Watanabe, T.: "Molecular characterization of the essential response regulator protein YycF in Bacillus subtilis"J.Molec.Microbiol.Biotechnol.. (印刷中). (2004)
[Publications] Sugiura, M.: "Identification and classification of two-component systems that affect rpoS expression in Escherichia coil."Biosci.Biotech.Biochem.. 67. 1612-1615 (2003)
[Publications] Hagiwara, D.: "Genome-wide analyses revealing a signaling network of the RcsC-YojN-RcsB phosphorelay system in Escherichia coil."J.Bacteriol.. 185. 5735-5746 (2003)
[Publications] Oshiro, T.: "A defect in a fatty acyl-CoA synthetase gene, lcf1l+, results in a decrease in viability after entry into the stationary phase in fission yeast."Mol.Genet.Genomics.. 269. 437-442 (2003)
[Publications] Nakamichi, N.: "Characterization of the Prr1 response regulator with special reference to sexual development in Schizosaccharomyces pombe."Biosci.Biotech.Biochem.. 67. 547-555 (2003)
[Publications] Sunohara, T.: "Ribosome stalling during translation elongation induces cleavage of mRNA being translated in Escherichia coil."J.Biol.Chem.. (印刷中). (2004)
[Publications] Morita, T.: "Accumulation of Glucose-6-phospnate or Fructose-6-phosphate Is Responsible for Destabilization of Glucose Transporter mRNA in Escherichia coli."J.Biol.Chem.. 278. 15608-15614 (2003)

2003 Fiscal Year Annual Research Report

大腸菌転写制御ネットワークの包括的解析

Principal Investigator

饗場 浩文 名古屋大学, 大学院・生命農学研究科, 助教授 (60211687)

Research Products

[Publications] Watanabe, T.: "Isolation and characterization of inhibitors of the essential histidine kinase, YycG in Bacillus subtilis and Staphylococcus aureus."J.Antibiotics. 56. 1045-1052 (2003)

[Publications] Eguchi, Y.: "Transcriptional regulation of drug efflux genes by EvgAS, a two-component system in Escherichia coli."Microbiology. 149. 2819-2828 (2003)

[Publications] Aketani, S.: "Role of a Nonnatural b-C-Nucleotide Unit in DNA as Template for DNA and RNA Synthesis and as Substrate for Nucleolytic Digestion."Eur.J.Phram.Sci.. 20. 43-51 (2003)

[Publications] Minagawa, S.: "Identification and molecular characterization of the Mg2+ stimulon of Escherichia coli."J.Bacteriol.. 185. 3696-3702 (2003)

[Publications] Gowrishankar, J.: "In Vitro Properties of RpoS (sigma(S)) Mutants of Escherichia coli with Postulated N-Terminal Subregion 1.1 or C-Terminal Region 4 Deleted."J.Bacteriol.. 185. 2673-2679 (2003)

[Publications] Yamamoto, K.: "Two different modes of transcription repression of the Escherichia coli acetate operon by Ic1R"Mol.Microbiol.. 47. 183-194 (2003)

[Publications] 中道範人: "分裂酵母における胞子形成制御するHis-Aspリン酸リレー情報伝達"日本農芸化学会誌. 78. 50-51 (2004)

[Publications] Yamamoto, Y.: "SsrA-mediated trans-translation plays a role in mRNA quality control by facilitating degradation of truncated mRNAs."RNA. 9. 408-418 (2003)

[Publications] EI-Kazzaz, W.: "Metabolic block at early stages of glycolytic pathway activates the Rcs phosphorelay system via increased synthesis of dTDP-glucose in Escherichia coli."Mol.Microbiol.. 51. 1117-1128 (2004)

[Publications] Sunohara, T.: "Nascent peptide-mediated ribosome stalling at a stop codon induces mRNA cleavage resulting in nonstop mRNA that is recognized by tmRNA."RNA. 10. 378-386 (2004)

[Publications] Eguchi, Y.: "A signal transduction cascade between EvgA/EvgS and PhoP/PhoQ two-component systems of Escherichia coli."J.Bacteriol.. (印刷中). (2004)

[Publications] Watanabe, T.: "Molecular characterization of the essential response regulator protein YycF in Bacillus subtilis"J.Molec.Microbiol.Biotechnol.. (印刷中). (2004)

[Publications] Sugiura, M.: "Identification and classification of two-component systems that affect rpoS expression in Escherichia coil."Biosci.Biotech.Biochem.. 67. 1612-1615 (2003)

[Publications] Hagiwara, D.: "Genome-wide analyses revealing a signaling network of the RcsC-YojN-RcsB phosphorelay system in Escherichia coil."J.Bacteriol.. 185. 5735-5746 (2003)

[Publications] Oshiro, T.: "A defect in a fatty acyl-CoA synthetase gene, lcf1l+, results in a decrease in viability after entry into the stationary phase in fission yeast."Mol.Genet.Genomics.. 269. 437-442 (2003)

[Publications] Nakamichi, N.: "Characterization of the Prr1 response regulator with special reference to sexual development in Schizosaccharomyces pombe."Biosci.Biotech.Biochem.. 67. 547-555 (2003)

[Publications] Sunohara, T.: "Ribosome stalling during translation elongation induces cleavage of mRNA being translated in Escherichia coil."J.Biol.Chem.. (印刷中). (2004)

[Publications] Morita, T.: "Accumulation of Glucose-6-phospnate or Fructose-6-phosphate Is Responsible for Destabilization of Glucose Transporter mRNA in Escherichia coli."J.Biol.Chem.. 278. 15608-15614 (2003)

饗場浩文名古屋大学, 大学院・生命農学研究科, 助教授 (60211687)