Functional analysis of Escherichia coli RNase G and its application for construction of protein expression system

大肠杆菌RNase G功能分析及其在蛋白表达系统构建中的应用

• 批准号: 15380059
• 负责人: WACHI Masaaki
• 金额: $ 9.28万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380059/
• 关键词: RNase G; RNase E; Hfq; FtsZ; stress protection; RNA metabolism; adhE; アルコールデヒドロゲナーゼ; バリン; ピルビン酸; 糖代謝; Gad; 酸耐性; エノラーゼ; cra; Dps; 酸化ストレス

In order to elucidate the regulatory mechanism of mRNA stability by the Escherichia coli endoribonuclease RNase G, action of RNase G on adhE mRNA was investigated. As a result, it was revealed that adhE mRNA cleaved by RNase G at -17 was rapidly degraded while one cleaved by RNase III at -31 was translated. It was found that RNase G mutant strains overproduced pyruvate because of increased stability of mRNAs encoding glycolysis enzymes. According to this finding, a new method for fermentative production of pyruvate and valine was established. It was found that processing of the ftsZ mRNA encoding the essential cell division protein FtsZ was an essential reaction of RNase E required for cell viability. In the RNase E mutant cells, translation of the unprocessed ftsZ mRNA was inhibited by the RNA-binding protein Hfq. For these results, it is suggested that mRNA stability control by endoribonucleases is widely involved in the regulatory mechanism of gene expression in E.coli.

为了阐明大肠杆菌核糖核酸内切酶RNase G对mRNA稳定性的调节机制，研究了RNase G对adhE mRNA的作用。结果显示，由RNase G在-17切割的adhE mRNA被快速降解，而由RNase III在-31切割的adhE mRNA被翻译。发现RNase G突变菌株过量产生丙酮酸，因为编码糖酵解酶的mRNA的稳定性增加。根据这一发现，建立了发酵生产丙酮酸和缬氨酸的新方法。发现编码必需的细胞分裂蛋白FtsZ的ftsZ mRNA的加工是细胞活力所需的RNase E的必需反应。在RNase E突变细胞中，未加工的ftsZ mRNA的翻译被RNA结合蛋白Hfq抑制。对于这些结果，它是建议，mRNA的稳定性控制由内切核糖核酸酶广泛参与大肠杆菌中的基因表达的调节机制。

项目成果

Isolation of a new antibiotic, alaremaycin, structurally related to 5-aminolevulinic acid from Streptomyces sp.A012304

从链霉菌属 sp.A012304 中分离出一种新抗生素阿拉霉素，其结构与 5-氨基乙酰丙酸相关

• 发表时间: 2005
• 期刊: Biosci.Biotechnol.Biochem. 69・9
• 作者: Awa;Y.
• 通讯作者: Y.

Takabumi Inagawa: "RNase ES of Streptomyces coelicolor A3(2) can complement the rne and rng mutations in Escherichia coli"Biosci.Biotechnol.Biochem.. 67. 1767-1771 (2003)

Takabumi Inakawa：“天蓝色链霉菌 A3(2) 的 RNase ES 可以补充大肠杆菌中的 rne 和 rng 突变”Biosci.Biotechnol.Biochem.. 67. 1767-1771 (2003)

A Corynebacterium glutamicum rnhA recG double mutant showing lysozyme-sensitivity, temperature-sensitivity growth, and UV-sensitivity.

谷氨酸棒杆菌 rnhA recG 双突变体表现出溶菌酶敏感性、温度敏感性生长和紫外线敏感性。

• 发表时间: 2003
• 期刊: Biosci.Biotechnol.Biochem. 67
• 作者: Hiwasawa;T.;Kumagai;Y.;Nagai;K.;Wachi;M.
• 通讯作者: M.

SulA-independent filamentation of Escherichia coli during growth after release from high hydrostatic pressure treatment

• DOI: 10.1007/s00253-003-1465-6
• 发表时间: 2004-04-01
• 期刊: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
• 影响因子: 5
• 作者: Kawarai, T;Wachi, M;Yamasaki, M
• 通讯作者: Yamasaki, M

Anucleate cell blue assay : a useful tool for identifying novel type II topoisomerase inhibitors

无核细胞蓝色测定：鉴定新型 II 型拓扑异构酶抑制剂的有用工具

• 发表时间: 2006
• 期刊: Antimicrob.Agents Chemother. 50・1
• 作者: Oyamada;Y.
• 通讯作者: Y.