1989 Fiscal Year Annual Research Report

ゲノム編制の全体像

Research Project

Project/Area Number	01655001
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) 井口八郎京都大学, 理学部, 助教授 (20028195) 伊藤維昭京都大学, ウイルス研究所, 教授 (90027334) 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	大腸菌ゲノムの編制 / 転写・翻訳制御因子 / tRNA遺伝子 / 分泌系内膜構成遺伝子 / 解糖系酵素遺伝子 / プリン生合成遺伝子 / IS因子とゲノムの再編 / 高度反復配列REP
Research Abstract	大腸菌ゲノム編成の全体像を明らかにするため、細胞活動の様々なレベルで重要な働きを担っている遺伝子、即ち、転写・翻訳系に関わるntrA、nusA、prfBなどの遺伝子、全tRNA遺伝子、分泌系内膜構成遺伝子、解糖系酵素遺伝子、プリン生合成遺伝子、DNA組換えや変換に関与する遺伝子の編制とゲノムの編成と再編成に関わる挿入因子ISや高度反復配列REPのゲノム上での分布と機能の解析を行った。まず、大腸菌tRNA遺伝子を新たに19個同定した。この結果、大腸菌tRNA遺伝子数は全部で79個存在し、これらは46種のtRNAに対応していて、41転写単位を構成していることが明らかとなった。次いで、転写・翻訳カスケード系に作用するnusAの変異に対するサプレッサー遺伝子を分離し、それがRNAポリメラーゼβ'サブユニットの変異であることを明らかにした。また分泌系内膜構成遺伝子編制の研究において、secY遺伝子産物中にリーダーペプチダーゼ認識部位の存在を見いだし、さらに、secY変異を抑制できるssyB、ssyDのクローニングを行った。解糖系酵素ホスホエノールピルビン酸カルボキシラーゼ遺伝子の部位突然変異法を用いて調べた結果、触媒活性部位と調節部位の領域に区別することができた。新たに5種のプリン生合成遺伝子、purE、purK、purH、purD、purLの塩基配列を決定して、その編制像を明らかにした、一方、大腸菌のゲノム編成と再編成の研究においては、各種のIS因子の内、IS1__ー、IS2__ー、IS3__ー、IS5__ー、IS<30>___ーの染色体上の全位置を決定するとともに、IS2__ーとIS3__ーのHfr形成との関係や、IS1__ーによるargF遺伝子の転移機構に関する新たな知見を得た。また、REP配列が大腸菌遺伝子組換えのホットスポットであることを示唆する結果を得、この配列の機能の一端を明らかにすることも成功した。

Research Products

(30 results)

All Other

All Publications (30 results)

[Publications] Watanabe,W.: "Identification and sequence analysis of Escherichia coli purE and purK genes encoding 5′-phosphoribosy1-5-amino-4-imidazole carboxy1-asc for do novo purine biosynthesis" J.Bacteriology. 171. 198-204 (1989)
[Publications] Sampei,G.: "The organization of the purL gene encoding 5′-phosphoribosylglycin amide amidotransferase of Escherichia coli" J.Biol.Chem.264. 21230-21238 (1989)
[Publications] Aiba,A.: "Nucleotide sequence analysis of genes purH and purD involved in the de novo purine nucleotide biosynthesis of Escherichia coli" J.Biol.Chem.264. 21239-21246 (1989)
[Publications] Hama,C.: "Organization of the replication control region of plasmid ColIb-P9" J.Bacteriology. 172. (1990)
[Publications] Shiba,K.: "Posttranscriptional control of repZ gene expression of the ColIb-P9 plasmid by a small RNA" J.Bacteriology. 172. (1990)
[Publications] Inouye,S.: "Cloning and sequence analysis of the ntrA(rpoN)gene of Pseudomonas putida" Gene.
[Publications] Inouye,S.: "Flagellation of Pseudomonas putida is dependent on the nerA(rpoN)gene" Mol.Gen.Genet.
[Publications] Inada,T.: "conditionally lethal amber mutations in the leader peptidase gene of Escherichia coli" J.Bacteriology. 171. 585-587 (1989)
[Publications] Nakamura,Y.: "Murine monoclonal antibodies which recognize active sites of Escherichia coli nusA protein and epitope mapping by gene fusion" Gene. 80. 13-19 (1989)
[Publications] Roesser,J.R.: "Regulation of basal level expression of the tryptophan operon of Escherichia coli" J.Biol.Chem.264. 12284-12288 (1989)
[Publications] Inada,T.: "Temperature-sensitive lethal mutant of Era,a G protein in Escherichia coli" J.Bacteriology. 171. 5017-5024 (1989)
[Publications] Bardwell,J.C.A.: "RNase III operon expression is autoregulated by mRNA processing" EMBO J.8. 3401-3407 (1989)
[Publications] Kawakami,K.: "Isolation and characterization of herC,a mutation of Escherichia coli affecting maintenance of ColEl" Mol.Gen.Genet.219. 333-340 (1989)
[Publications] Ozeki,H.: "The chloroplast genome of plants;a unique origin" Genome. 31. 169-174 (1989)
[Publications] Kondo,K.: "The transposable element Tn3 promotes general recombination at the neighboring regions" Jpn.J.Genetics. 64. 417-434 (1989)
[Publications] Komine,Y.: "Genomic organization and physical mapping of the transfer RNA genes in Escherichia coli" J.Mol.Biol.211. (1990)
[Publications] Akiyama,Y.: "Export of Escherichia coli alkaline phosphatase attached to an integral membrane protein,SecY" J.Biol.Chem.264. 437-442 (1989)
[Publications] Ito,K.: "Temperature-sensitive mutants of Escherichia coli;inhibition of protein export at the permissive temperature" J.Bacteriology. 171. 1742-1743 (1989)
[Publications] Ueguchi,C.: "The secY-ipmJ region of the spc ribosomal protein operon in Escherichia coli;structural alterations affecting secY expression" Mol.Gen.Genet.217. 1-5 (1989)
[Publications] Lill,R.: "SecA protein hydrolyzes ATP and is essential component of the protein translocation ATPase of Escherichia coli" EMBO J.8. 961-966 (1989)
[Publications] Kusukawa,N.: "Effects of mutations in heat-shock genes groES and groEL on protein export in Escherichia coli" EMBO J.8. 3571-3521 (1989)
[Publications] Taura,T.: "Transient shut off of Escherichia coli heat-shock protein synthesis upon temperature shift down" Biochem.Biophys.Res.Commun.163. 438-443 (1989)
[Publications] Ito,K.: "Structure,function and biogenesis of SecY,an integral membrane protein involved in protein export" J.Bioenerg.Biomemb.
[Publications] Inoue,M.: "First crystallization of a phosphoenolpyruvate carboxylase from Escherichia coli" J.Mol.Biol.208. 509-510 (1989)
[Publications] Umeda,M.: "Mapping of insertion elements IS1,IS2,and IS3 on the E.coli chromosome.Role of insertion elements in formation of Hfrs and F-prime factors and in rearrangement" J.Mol.Biol.208. 601-614 (1989)
[Publications] Sekine,K.: "Frameshifting is required for expression of IS1 transposase" Proc.Natl.Acad.Sci USA. 86. 4609-4613 (1989)
[Publications] Yasue,H.: "The oncogenicity of avian adenoviruses.IV.Confirmatory evidence for recombination between viral and cellular DNA sequences and repetition of the recombinant in cells of a tumore line" Virology. 169. 447-451 (1989)
[Publications] Umeda,M.: "Mapping of an insertion element IS5 in the E.coli K-12 chromosome:Role of IS5 in rearrangement of the bacterial chromosome" J.Mol.Biol.(1990)
[Publications] Umeda,M.: "Mapping of insertion element IS30 on the Escherichia coli K-12 chromosome" Mol.Gen.Genet.
[Publications] Tenzen,T.: "Site-specific transposition of insertion element IS30" J.Bacteriology.

1989 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] Watanabe,W.: "Identification and sequence analysis of Escherichia coli purE and purK genes encoding 5′-phosphoribosy1-5-amino-4-imidazole carboxy1-asc for do novo purine biosynthesis" J.Bacteriology. 171. 198-204 (1989)

[Publications] Sampei,G.: "The organization of the purL gene encoding 5′-phosphoribosylglycin amide amidotransferase of Escherichia coli" J.Biol.Chem.264. 21230-21238 (1989)

[Publications] Aiba,A.: "Nucleotide sequence analysis of genes purH and purD involved in the de novo purine nucleotide biosynthesis of Escherichia coli" J.Biol.Chem.264. 21239-21246 (1989)

[Publications] Hama,C.: "Organization of the replication control region of plasmid ColIb-P9" J.Bacteriology. 172. (1990)

[Publications] Shiba,K.: "Posttranscriptional control of repZ gene expression of the ColIb-P9 plasmid by a small RNA" J.Bacteriology. 172. (1990)

[Publications] Inouye,S.: "Cloning and sequence analysis of the ntrA(rpoN)gene of Pseudomonas putida" Gene.

[Publications] Inouye,S.: "Flagellation of Pseudomonas putida is dependent on the nerA(rpoN)gene" Mol.Gen.Genet.

[Publications] Inada,T.: "conditionally lethal amber mutations in the leader peptidase gene of Escherichia coli" J.Bacteriology. 171. 585-587 (1989)

[Publications] Nakamura,Y.: "Murine monoclonal antibodies which recognize active sites of Escherichia coli nusA protein and epitope mapping by gene fusion" Gene. 80. 13-19 (1989)

[Publications] Roesser,J.R.: "Regulation of basal level expression of the tryptophan operon of Escherichia coli" J.Biol.Chem.264. 12284-12288 (1989)

[Publications] Inada,T.: "Temperature-sensitive lethal mutant of Era,a G protein in Escherichia coli" J.Bacteriology. 171. 5017-5024 (1989)

[Publications] Bardwell,J.C.A.: "RNase III operon expression is autoregulated by mRNA processing" EMBO J.8. 3401-3407 (1989)

[Publications] Kawakami,K.: "Isolation and characterization of herC,a mutation of Escherichia coli affecting maintenance of ColEl" Mol.Gen.Genet.219. 333-340 (1989)

[Publications] Ozeki,H.: "The chloroplast genome of plants;a unique origin" Genome. 31. 169-174 (1989)

[Publications] Kondo,K.: "The transposable element Tn3 promotes general recombination at the neighboring regions" Jpn.J.Genetics. 64. 417-434 (1989)

[Publications] Komine,Y.: "Genomic organization and physical mapping of the transfer RNA genes in Escherichia coli" J.Mol.Biol.211. (1990)

[Publications] Akiyama,Y.: "Export of Escherichia coli alkaline phosphatase attached to an integral membrane protein,SecY" J.Biol.Chem.264. 437-442 (1989)

[Publications] Ito,K.: "Temperature-sensitive mutants of Escherichia coli;inhibition of protein export at the permissive temperature" J.Bacteriology. 171. 1742-1743 (1989)

[Publications] Ueguchi,C.: "The secY-ipmJ region of the spc ribosomal protein operon in Escherichia coli;structural alterations affecting secY expression" Mol.Gen.Genet.217. 1-5 (1989)

[Publications] Lill,R.: "SecA protein hydrolyzes ATP and is essential component of the protein translocation ATPase of Escherichia coli" EMBO J.8. 961-966 (1989)

[Publications] Kusukawa,N.: "Effects of mutations in heat-shock genes groES and groEL on protein export in Escherichia coli" EMBO J.8. 3571-3521 (1989)

[Publications] Taura,T.: "Transient shut off of Escherichia coli heat-shock protein synthesis upon temperature shift down" Biochem.Biophys.Res.Commun.163. 438-443 (1989)

[Publications] Ito,K.: "Structure,function and biogenesis of SecY,an integral membrane protein involved in protein export" J.Bioenerg.Biomemb.

[Publications] Inoue,M.: "First crystallization of a phosphoenolpyruvate carboxylase from Escherichia coli" J.Mol.Biol.208. 509-510 (1989)

[Publications] Umeda,M.: "Mapping of insertion elements IS1,IS2,and IS3 on the E.coli chromosome.Role of insertion elements in formation of Hfrs and F-prime factors and in rearrangement" J.Mol.Biol.208. 601-614 (1989)

[Publications] Sekine,K.: "Frameshifting is required for expression of IS1 transposase" Proc.Natl.Acad.Sci USA. 86. 4609-4613 (1989)

[Publications] Yasue,H.: "The oncogenicity of avian adenoviruses.IV.Confirmatory evidence for recombination between viral and cellular DNA sequences and repetition of the recombinant in cells of a tumore line" Virology. 169. 447-451 (1989)

[Publications] Umeda,M.: "Mapping of an insertion element IS5 in the E.coli K-12 chromosome:Role of IS5 in rearrangement of the bacterial chromosome" J.Mol.Biol.(1990)

[Publications] Umeda,M.: "Mapping of insertion element IS30 on the Escherichia coli K-12 chromosome" Mol.Gen.Genet.

[Publications] Tenzen,T.: "Site-specific transposition of insertion element IS30" J.Bacteriology.

溝渕潔東京大学, 理学部, 助教授 (00092346)