1989 年度実績報告書

ゲノム編制の全体像

研究課題

研究課題/領域番号	01655001
研究機関	東京大学
研究代表者	溝渕潔東京大学, 理学部, 助教授 (00092346)
研究分担者	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)
キーワード	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
研究概要	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。

研究成果

(30件)

すべてその他

すべて文献書誌 (30件)

[文献書誌] 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)
[文献書誌] Sampei,G.: "The organization of the purL gene encoding 5′-phosphoribosylglycin amide amidotransferase of Escherichia coli" J.Biol.Chem.264. 21230-21238 (1989)
[文献書誌] 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)
[文献書誌] Hama,C.: "Organization of the replication control region of plasmid ColIb-P9" J.Bacteriology. 172. (1990)
[文献書誌] Shiba,K.: "Posttranscriptional control of repZ gene expression of the ColIb-P9 plasmid by a small RNA" J.Bacteriology. 172. (1990)
[文献書誌] Inouye,S.: "Cloning and sequence analysis of the ntrA(rpoN)gene of Pseudomonas putida" Gene.
[文献書誌] Inouye,S.: "Flagellation of Pseudomonas putida is dependent on the nerA(rpoN)gene" Mol.Gen.Genet.
[文献書誌] Inada,T.: "conditionally lethal amber mutations in the leader peptidase gene of Escherichia coli" J.Bacteriology. 171. 585-587 (1989)
[文献書誌] 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)
[文献書誌] Roesser,J.R.: "Regulation of basal level expression of the tryptophan operon of Escherichia coli" J.Biol.Chem.264. 12284-12288 (1989)
[文献書誌] Inada,T.: "Temperature-sensitive lethal mutant of Era,a G protein in Escherichia coli" J.Bacteriology. 171. 5017-5024 (1989)
[文献書誌] Bardwell,J.C.A.: "RNase III operon expression is autoregulated by mRNA processing" EMBO J.8. 3401-3407 (1989)
[文献書誌] Kawakami,K.: "Isolation and characterization of herC,a mutation of Escherichia coli affecting maintenance of ColEl" Mol.Gen.Genet.219. 333-340 (1989)
[文献書誌] Ozeki,H.: "The chloroplast genome of plants;a unique origin" Genome. 31. 169-174 (1989)
[文献書誌] Kondo,K.: "The transposable element Tn3 promotes general recombination at the neighboring regions" Jpn.J.Genetics. 64. 417-434 (1989)
[文献書誌] Komine,Y.: "Genomic organization and physical mapping of the transfer RNA genes in Escherichia coli" J.Mol.Biol.211. (1990)
[文献書誌] Akiyama,Y.: "Export of Escherichia coli alkaline phosphatase attached to an integral membrane protein,SecY" J.Biol.Chem.264. 437-442 (1989)
[文献書誌] Ito,K.: "Temperature-sensitive mutants of Escherichia coli;inhibition of protein export at the permissive temperature" J.Bacteriology. 171. 1742-1743 (1989)
[文献書誌] 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)
[文献書誌] Lill,R.: "SecA protein hydrolyzes ATP and is essential component of the protein translocation ATPase of Escherichia coli" EMBO J.8. 961-966 (1989)
[文献書誌] Kusukawa,N.: "Effects of mutations in heat-shock genes groES and groEL on protein export in Escherichia coli" EMBO J.8. 3571-3521 (1989)
[文献書誌] Taura,T.: "Transient shut off of Escherichia coli heat-shock protein synthesis upon temperature shift down" Biochem.Biophys.Res.Commun.163. 438-443 (1989)
[文献書誌] Ito,K.: "Structure,function and biogenesis of SecY,an integral membrane protein involved in protein export" J.Bioenerg.Biomemb.
[文献書誌] Inoue,M.: "First crystallization of a phosphoenolpyruvate carboxylase from Escherichia coli" J.Mol.Biol.208. 509-510 (1989)
[文献書誌] 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)
[文献書誌] Sekine,K.: "Frameshifting is required for expression of IS1 transposase" Proc.Natl.Acad.Sci USA. 86. 4609-4613 (1989)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Umeda,M.: "Mapping of insertion element IS30 on the Escherichia coli K-12 chromosome" Mol.Gen.Genet.
[文献書誌] Tenzen,T.: "Site-specific transposition of insertion element IS30" J.Bacteriology.

1989 年度 実績報告書

ゲノム編制の全体像

研究代表者

溝渕 潔 東京大学, 理学部, 助教授 (00092346)

研究成果

[文献書誌] 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)

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

[文献書誌] 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)

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

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

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

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

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

[文献書誌] 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)

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

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

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

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

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

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

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

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

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

[文献書誌] 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)

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

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

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

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

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

[文献書誌] 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)

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

[文献書誌] 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)

[文献書誌] 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)

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

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

1989 年度実績報告書

溝渕潔東京大学, 理学部, 助教授 (00092346)