Study on the transcriptional control of flagellar regulon in Salmonella typhimurium.

鼠伤寒沙门氏菌鞭毛调节子转录调控的研究。

• 批准号: 63540500
• 负责人: KUTSUKAKE Kazuhiro
• 金额: $ 1.22万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63540500/
• 关键词: Bacterial flagella; Flagellar regulon; Gene expression; Transcriptional regulation; Regulatory protein; Derepressed mutant; In vitro transcription; Protein purification; べん毛レギュロン

1. An overall structure of flagellar regulon of Salmonella has been established. Based on the transcriptional hierarchy. flagellar operons can be divided into three classes (Class I. Class II, and Class III). The transcription of the class III operons is controlled positively by the gene product of fliA which belongs to class II and negatively by the product of a newly identified gene, rflB, which belongs to class III.2. The fliA gene was cloned and sequenced. The deduced animo acid sequence of the FliA protein was revealed to share a great deal of homology with those of the known sigma factors of RNA polymerase. The purified FliA protein had an activity to promote specifically the transcription from the promoter of flagellar operons when added to the in vitro transcription system containing RNA polymerase core enzyme. This indicates that the FliA protein is an alternative sigma factor specific for flagellar regulon.3. The rflB gene was cloned and sequenced. The purified RflB protein had an activity to repress specifically the transcription from the promoter of flagellar operons when added to the in vitro transcription system containing FliA-RNA polymerase.4. A number of derepressed mutants of the class III operons were isolated. Some of them were found to have mutations in the fliA gene, which suggests that there may be a molecular interaction between the FliA and the RflB proteins. This was proved in vitro using the purified FliA and RflB proteins.5. Expression of the class II operons was shown to be enhanced in the rflB mutants. This enhancement disappeared when the fliA defect was introduced into the same strains. Therefore, it has been concluded that the activity of the whole flagellar regulon is controlled positively and negatively by the FliA and the RflB proteins. respectively.

1.建立了沙门氏菌鞭毛调节子的整体结构。基于转录层次结构。鞭毛操纵子可分为三类（I类、II类和III类）。 III类操纵子的转录由属于II类的fliA的基因产物正向控制，并且由属于III.2类的新鉴定的基因rflB的产物负向控制。克隆并测序了fliA基因。推导的 FliA 蛋白氨基酸序列与已知的 RNA 聚合酶 sigma 因子的氨基酸序列具有很大的同源性。当将纯化的FliA蛋白添加到含有RNA聚合酶核心酶的体外转录系统时，具有特异性促进鞭毛操纵子启动子转录的活性。这表明FliA蛋白是鞭毛调节子特异的另一种西格玛因子。3． rflB 基因被克隆并测序。纯化的RflB蛋白加入到含有FliA-RNA聚合酶的体外转录系统中，具有特异性抑制鞭毛操纵子启动子转录的活性。 4．分离出许多III类操纵子的去抑制突变体。其中一些被发现在 fliA 基因中存在突变，这表明 FliA 和 RflB 蛋白之间可能存在分子相互作用。使用纯化的FliA和RflB蛋白在体外证明了这一点。5． rflB 突变体中 II 类操纵子的表达得到增强。当将fliA缺陷引入相同菌株时，这种增强消失了。因此，可以得出结论，整个鞭毛调节子的活性受FliA和RflB蛋白的正向和负向控制。分别。

项目成果

沓掛和弘: "微生物学ハンドブック" 石川辰夫、駒形和男、杉山純多、田中健治、堀寛、水島昭二、柳田友道 編 丸善株式会社, (1990)

Kazuhiro Kutsukake：“微生物学手册”，石川龙夫、驹形一夫、杉山顺太、田中健二、堀浩、水岛正司、柳田智通编，丸善株式会社（1990 年）

Kutsukake, K., Ohya, Y., Yamaguchi, S., Iino, T.: "Operon structure of flagellar genes in Salmonella typhimurium." Mol. Gen. Genet. 214 11-15, 1988.

Kutsukake, K.、Ohya, Y.、Yamaguchi, S.、Iino, T.：“鼠伤寒沙门氏菌鞭毛基因的操纵子结构。”

Iino,T.;Komeda,Y.;Kutsukake,K.;Macnab,R.M.;Matsumura,P.;Parkinson,J.S.;Simon,M.I.;Yamaguchi,S.: Microbiol.Rev.52. 533-535 (1988)

Iino，T.；Komeda，Y.；Kutsukake，K.；Macnab，R.M.；Matsumura，P.；Parkinson，J.S.；Simon，M.I.；Yamaguchi，S.：微生物学Rev.52。

Kutsukake,K.,Ohya,Y.,Yamaguchi,S.,Iino.t.: "Operon structure of flagellar genes in Salmonella typhimurium." Molecular and General Genetics. 214. 11-15 (1988)

Kutsukake，K.，Ohya，Y.，Yamaguchi，S.，Iino.t.：“鼠伤寒沙门氏菌鞭毛基因的操纵子结构。”

Wada, M., Kutsukake, K., Komano, T., Imamoto, F., Kano, Y.: "Participation of the hup gene product in site-specific DNA inversion in Escherichia coli." Gene 76 345-352, 1989.

Wada, M.、Kutsukake, K.、Komano, T.、Imamoto, F.、Kano, Y.：“hup 基因产物参与大肠杆菌中的位点特异性 DNA 倒转。”