1991 Fiscal Year Annual Research Report

ゲノム編制の全体像

Research Project

Project/Area Number	03221101
Research Institution	The University of Tokyo
Principal Investigator	溝渕潔東京大学, 理学部, 助教授 (00092346)
Co-Investigator(Kenkyū-buntansha)	大坪栄一東京大学, 応用微生物研究所, 教授 (10158800) 伊藤維昭京都大学, ウイルス研究所, 教授 (90027334) 井口八郎京都大学, 理学部, 助教授 (20028195) 中村義一東京大学, 医科学研究所, 助教授 (40114590) 中沢淳山口大学, 医学部, 教授 (90025594)
Keywords	大腸菌ゲノム編制 / 転写開始因子 / アミノアシルtRNA合成酵素 / 低分子RNA10Sa / 分泌系内膜構成遺伝子 / ヘム合成遺伝子 / IS3挿入因子 / 反復配列
Research Abstract	大腸菌ゲノム(470万塩基対)は約4000遺伝子から構成されている。現在まで約1500遺伝子が検出・同定され、その内1000種の遺伝子について塩基配列が決定された。本研究では大腸菌ゲノム編制の全体像を明らかにするため、転写開始と転写・翻訳カスケ-ド反応に関わる遺伝子群、低分子RNA10Sa遺伝子、蛋白質分泌系内膜構成遺伝子群、解糖系酵素遺伝子群、ヘム合成遺伝子群、グルクロン酸、ガラクツロン酸代謝遺伝子群、細胞分裂に関与する遺伝子群などの編制と機能を明らかにするとともに、ゲノムの再編に関わる各種の挿入配列(IS因子)のゲノム上での分布と構造を取り上げて解析を進めてきた。本研究で得られた主な結果は以下の通りである。(1)大腸菌転写開始因子の1つであるσ^<54>をコ-ドするntrA遺伝子周辺の塩基配列を解析したところ、その編制はKlebsiella pneumoniaeをはじめ様々な細菌でよく保存されていることが明らかとなった。(2)翻訳反応に関わるリジンtRNA合成酵素遺伝子lysUの発現は、ロイシン代謝の制御に関わるLrp蛋白質によって調節されていることを新たに見いだした。また、このことと関連して、(3)細胞分裂の頻度を決定している遺伝子cfcAの構造を解析したところ、グリシンtRNA合成酵素であることが明らかとなった。これらの結果はアミノアシルtRNA合成酵素の新たな機能面での問題を提起している。(4)ヘム合成に関与する8遺伝子(hemAーhemH)、グルクロン酸、ガラクツロン酸代謝に関与する8遺伝子、低分子RNA10Sa遺伝子、分泌系においてペリプラズム蛋白質のSーS結合形成に必須のppfA遺伝子の同定とその編制像を明らかにした。(5)ゲノム再編に関与するIS3因子はゲノム上4箇所に挿入されているが、それらはすべて同じ配列を持ち、2つのIS3近傍で欠失が生じていることが明らかとなった。(6)また、これらの研究過程において、123塩基の新しい反復配列を発見した。

Research Products

(23 results)

All Other

All Publications (23 results)

[Publications] K.Asano: "Positive and negative regulations of plasmid ColIb-P9 repZ gene expression at the translational level" J.Biol.Chem.266. 3774-3781 (1991)
[Publications] K.Asano: "An induced mRNA secondary structure enhances repZ translation in plasmid ColIb-P9" J.Biol.Chem.266. 424549-24556 (1991)
[Publications] Y.Yonetani: "Isolation and characterization of an adenine sensitive mutant from Escherichia coli K-12" J.Bacteriol.174. (1992)
[Publications] K.Matsumura: "Cloning and sequencing of the uxa and uxu genes involved in glucuronate and galacturonate metabolism in Eecherichia coli" J.Bacteriol.174. (1992)
[Publications] V.Gomada: "Analysis of upstream regulatory sequence for activation of the regulatory gene xyls in xylene metabolism directed by the TOL plasmid of Pseudomonas putida" Mol.Gen.Genet.
[Publications] K.Ito: "Genetic interaction between subunit of RNA polymerase and arginine-rich domain of Escherichia coli nusA protein" J.Bacteriol.173. 1492-1501 (1991)
[Publications] O.Mikumi: "Sequence and functional analysis of mutations in the gene encoding peptide-chain-release factor 2 of Escherichia coli" Biochimie. 73. 1509-1517 (1991)
[Publications] K.Nakahigashi: "Photosensitivity of a protoporphyrin-accumulating,light-sensitive mutant (visA) of Escherichia coli K-12" Proc.Natl.Acad.Sci.USA. 88. 10520-10524 (1991)
[Publications] K.Miyamoto: "Isolation and characterization of visible light-sensitive mutants of Escherichia coli K-12" J.Mol.Biol.219. 393-398 (1991)
[Publications] Y.Komine: "Precise mapping of the rnpB gene encoding the RNA component of RNaseP in Escherichia coli K-12" J.Bacteriol.173. 1813-1816 (1991)
[Publications] Y.Komine: "Physical map locations of the genes that encode small stable RNAs in Escherichia coli K-12" J.Bacteriol.173. 5252 (1991)
[Publications] M.Sakaguchi: "Yeast gene which suppresses the defect in protein export of a secY mutant of E.coli" J.Biochem.109. 799-802 (1991)
[Publications] T.Taura: "Does protein secretion activity vary during the cell cycle of Escherichia coli" J.Biochem.109. 811-815 (1991)
[Publications] K.Ito: "In vivo analysis of integration of membrane proteins in Escherichia coli" Mol.Microbiol.5. 2243-2253 (1991)
[Publications] S.Kamitani: "Identification and characterization of an Escherichia coli gene required for the formation of correctly folded alkaline phosphatase" EMBO J.11. 57-62 (1991)
[Publications] C.Ueguchi: "Multicopy suppression:an approach to understanding intracellular functioning of the protein export system" J.Bacteriol.174. (1992)
[Publications] T.shimoike: "SecY variants that interfere with Escherichia coli protein export in the presence of normal secY" Mol.Microbiol.6. (1992)
[Publications] K.Terada: "Site-directed mutagenesis of the conserved histidine residue of phosphoenolpyruvate carboxylase" Eur.J.Biochem.202. 797-803 (1991)
[Publications] A.Nishimura: "Correlation of the pLC-plasmids to the physical map of Escherichia coli K-12" Microbiol.Rev.56. (1992)
[Publications] M.Umeda: "Four types of IS1 with difference in nucleotide sequences reside in the Escherichia coli K-12 chromosome" Gene. 98. 1-5 (1991)
[Publications] T.Tenzen: "Preferential transposition of an IS630-associated composite transposon to TA in the 5'-CTAG-3' sequence" J.Bacteriol.173. 6207-6212 (1991)
[Publications] A.Nishimura: "Control of cell growth and division" Springer-Verlag/Japan Sci.Soc.Press,Tokyo, 266 (1991)
[Publications] Y.Sekine: "New aspects of the genetics of molecular evolution" Springer-Verlag/Japan Sci.Soc.Press,Tokyo, 19 (1991)

1991 Fiscal Year Annual Research Report

ゲノム編制の全体像

Principal Investigator

溝渕 潔 東京大学, 理学部, 助教授 (00092346)

Research Products

[Publications] K.Asano: "Positive and negative regulations of plasmid ColIb-P9 repZ gene expression at the translational level" J.Biol.Chem.266. 3774-3781 (1991)

[Publications] K.Asano: "An induced mRNA secondary structure enhances repZ translation in plasmid ColIb-P9" J.Biol.Chem.266. 424549-24556 (1991)

[Publications] Y.Yonetani: "Isolation and characterization of an adenine sensitive mutant from Escherichia coli K-12" J.Bacteriol.174. (1992)

[Publications] K.Matsumura: "Cloning and sequencing of the uxa and uxu genes involved in glucuronate and galacturonate metabolism in Eecherichia coli" J.Bacteriol.174. (1992)

[Publications] V.Gomada: "Analysis of upstream regulatory sequence for activation of the regulatory gene xyls in xylene metabolism directed by the TOL plasmid of Pseudomonas putida" Mol.Gen.Genet.

[Publications] K.Ito: "Genetic interaction between subunit of RNA polymerase and arginine-rich domain of Escherichia coli nusA protein" J.Bacteriol.173. 1492-1501 (1991)

[Publications] O.Mikumi: "Sequence and functional analysis of mutations in the gene encoding peptide-chain-release factor 2 of Escherichia coli" Biochimie. 73. 1509-1517 (1991)

[Publications] K.Nakahigashi: "Photosensitivity of a protoporphyrin-accumulating,light-sensitive mutant (visA) of Escherichia coli K-12" Proc.Natl.Acad.Sci.USA. 88. 10520-10524 (1991)

[Publications] K.Miyamoto: "Isolation and characterization of visible light-sensitive mutants of Escherichia coli K-12" J.Mol.Biol.219. 393-398 (1991)

[Publications] Y.Komine: "Precise mapping of the rnpB gene encoding the RNA component of RNaseP in Escherichia coli K-12" J.Bacteriol.173. 1813-1816 (1991)

[Publications] Y.Komine: "Physical map locations of the genes that encode small stable RNAs in Escherichia coli K-12" J.Bacteriol.173. 5252 (1991)

[Publications] M.Sakaguchi: "Yeast gene which suppresses the defect in protein export of a secY mutant of E.coli" J.Biochem.109. 799-802 (1991)

[Publications] T.Taura: "Does protein secretion activity vary during the cell cycle of Escherichia coli" J.Biochem.109. 811-815 (1991)

[Publications] K.Ito: "In vivo analysis of integration of membrane proteins in Escherichia coli" Mol.Microbiol.5. 2243-2253 (1991)

[Publications] S.Kamitani: "Identification and characterization of an Escherichia coli gene required for the formation of correctly folded alkaline phosphatase" EMBO J.11. 57-62 (1991)

[Publications] C.Ueguchi: "Multicopy suppression:an approach to understanding intracellular functioning of the protein export system" J.Bacteriol.174. (1992)

[Publications] T.shimoike: "SecY variants that interfere with Escherichia coli protein export in the presence of normal secY" Mol.Microbiol.6. (1992)

[Publications] K.Terada: "Site-directed mutagenesis of the conserved histidine residue of phosphoenolpyruvate carboxylase" Eur.J.Biochem.202. 797-803 (1991)

[Publications] A.Nishimura: "Correlation of the pLC-plasmids to the physical map of Escherichia coli K-12" Microbiol.Rev.56. (1992)

[Publications] M.Umeda: "Four types of IS1 with difference in nucleotide sequences reside in the Escherichia coli K-12 chromosome" Gene. 98. 1-5 (1991)

[Publications] T.Tenzen: "Preferential transposition of an IS630-associated composite transposon to TA in the 5'-CTAG-3' sequence" J.Bacteriol.173. 6207-6212 (1991)

[Publications] A.Nishimura: "Control of cell growth and division" Springer-Verlag/Japan Sci.Soc.Press,Tokyo, 266 (1991)

[Publications] Y.Sekine: "New aspects of the genetics of molecular evolution" Springer-Verlag/Japan Sci.Soc.Press,Tokyo, 19 (1991)

溝渕潔東京大学, 理学部, 助教授 (00092346)