2018 Fiscal Year Final Research Report
Identification of the whole set of constitutive promoters in Escherichia coli
| Project/Area Number |
16K07195
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Genome biology
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| Research Institution | Meiji University (2017-2018)
Tokyo Institute of Technology (2016) |
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Principal Investigator |
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| Research Collaborator |
Ishihama Akira 法政大学, マイクロ・ナノテクノロジー研究所, 特任教授
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Keywords | ゲノム / 転写制御 / RNAポリメラーゼ / シグマ因子 / Genomic SELEX / 遺伝子発現 / 大腸菌 / constitutive promoter |
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| Outline of Final Research Achievements |
The promoter selectivity of Escherichia coli RNA polymerase (RNAP) is determined by the sigma subunit. The model prokaryote Escherichia coli K-12 contains seven species of the sigma subunit, each recognizing a specific set of promoters. For identification of the “constitutive promoters” that are recognized by each RNAP holoenzyme alone in the absence of other supporting factors, we have performed the genomic SELEX screening in vitro for their binding sites along the E. coli K-12 W3110 genome using each of the reconstituted RNAP holoenzymes and a collection of genome DNA segments of E. coli K-12. The whole set of constitutive promoters for each RNAP holoenzyme was then estimated based on the location of RNAP-binding sites. Five sigma factors, RpoD, RpoS, RpoD, RpoF, RpoE were succeeded to identify the constitutive promoters, and the list was published in original article. The list of constitutive promoters provides the fundamental catalogs for the promoter recognized in E.coli.
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| Free Research Field |
ゲノム転写制御
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| Academic Significance and Societal Importance of the Research Achievements |
大腸菌の持つ全７種類のシグマ因子のうち、RpoD, RpoS, RpoH, RpoF, RpoEの５つのシグマ因子について、RNAポリメラーゼホロ酵素単独で認識されるプロモーターの同定に成功した。また、それらの結果を統合し、ゲノム転写制御機構の全体像のモデルを提案した。そして、その成果を原著論文として発表した。これらの成果は、生物がゲノムにある遺伝子を利用する仕組みの本質的な理解に役立つと考えられる。また、本成果の情報は、モデル生物である大腸菌の研究コミュニティでも活用されることが期待される。
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