Characterization of trigger enzymes involved in the regulation and function of sRNAs in B. subtilis

枯草芽孢杆菌中参与 sRNA 调节和功能的触发酶的表征

• 批准号: 238802965
• 负责人: Privatdozentin Dr. Sabine Brantl
• 金额: --
• 依托单位: Lehrstuhl für Genetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/238802965?language=en
• 关键词: Characterization; trigger; enzymes; involved; regulation

So far, only a few so called trigger enzymes are known. These are metabolic enzymes that fulfill a second function in control of gene expression. Trigger enzymes known to date can e.g. bind DNA and act as transcription factors, affect the stability of RNA by binding it, modify proteins by e.g. phosphorylation or inhibit transcription factors by protein/protein interactions. Furthermore, compared to E. coli and other Gram-negative bacteria, not so much is known about sRNAs and their targest in Gram-positive bacteria, in particular B. subtilis. In the latter bacterium, only for five sRNAs targets have been identified, among them for the two sRNAs SR1 and SR4 investigated in our group.In this project we want toa) characterize a novel trigger enzyme, pyruvate carboxylase A (PycA) recently identified in our group as second regulator of the sRNA SR1, in great detail biochemically and concerning its biological role. Both the mechanism of action and the structural and biological prerequisites for its action should be elucidated.b) identify further targets of PycA and analyse the action of this trigger enzymes at the promoters of these target gens.c) demonstrate that glyceraldehyde-3-phosphate-dehydrogenase GapA from B. subtilis is also a trigger enzyme (either a RNase or a protein that interacts with an RNAse or components of the degradosome), whose function is modulated by the small peptide SR1P encoded by SR1d) identify further RNAs whose stability depends on GapA, and investigate the mechanism of action of GapA at these RNAs.

到目前为止，只有少数所谓的触发酶已知。这些代谢酶在控制基因表达中发挥第二种功能。迄今已知的触发酶可以例如结合DNA并充当转录因子，通过结合RNA影响RNA的稳定性，通过例如磷酸化修饰蛋白质或通过蛋白质/蛋白质相互作用抑制转录因子。此外，与E.大肠杆菌和其它革兰氏阴性菌中的sRNA，对于革兰氏阳性菌，特别是B中的sRNA及其靶点知之甚少。枯草芽孢杆菌在后一种细菌中，只鉴定了5种sRNA靶点，其中两种sRNA SR 1和SR 4是我们小组研究的。在这个项目中，我们想a）表征一种新的触发酶，丙酮酸羧化酶A（PycA），它是我们小组最近鉴定的sRNA SR 1的第二个调节因子，在生物化学和生物学作用方面非常详细。B）鉴定PycA的进一步靶点并分析该触发酶在这些靶基因的启动子处的作用。c）证明甘油醛-3-磷酸脱氢酶GapA来自B。枯草芽孢杆菌也是一种触发酶（RNA酶或与RNA酶或降解体组分相互作用的蛋白质），其功能由SR 1d编码的小肽SR 1 P调节）鉴定其稳定性取决于GapA的其他RNA，并研究GapA对这些RNA的作用机制。