Štěpánek Václav, Dostálová Hana, Busche Tobias, Blumenstein Jan, Grulich Michal, Plašil Lukáš, Rucká Lenka, Nešvera Jan, Pátek Miroslav
Institute of Microbiology of the CAS, v.v.i., Videňská 1083, CZ-14220 Prague 4, Czech Republic.
Center for Biotechnology, Bielefeld University, Universitätsstrasse 27, D-33615 Bielefeld, Germany.
FEMS Microbiol Lett. 2022 Feb 22;369(1). doi: 10.1093/femsle/fnac014.
The aim of this investigation was to discover the promoters that drive expression of the sig genes encoding sigma factors of RNA polymerase in Rhodococcus erythropolis CCM2595 and classify these promoters according to the sigma factors which control their activity. To analyze the regulation of major sigma factors, which control large regulons that also contain genes expressed under exponential growth and non-stressed conditions, we used the R. erythropolis CCM2595 culture, which grew rapidly in minimal medium. The transcriptional start sites (TSSs) of the genes sigA, sigB, sigD, sigE, sigG, sigH, sigJ, and sigK were detected by primary 5'-end-specific RNA sequencing. The promoters localized upstream of the detected TSSs were defined by their -35 and -10 elements, which were identical or closely similar to these sequences in the related species Corynebacterium glutamicum and Mycobacterium tuberculosis. Regulation of the promoter activities by different sigma factors was demonstrated by two independent techniques (in vivo and in vitro). All analyzed sig genes encoding the sigma factors with extracytoplasmic function (ECF) were found to be also driven from additional housekeeping promoters. Based on the classification of the sig gene promoters, a model of the basic sigma transcriptional regulatory network in R. erythropolis was designed.
本研究的目的是发现驱动红平红球菌CCM2595中编码RNA聚合酶σ因子的sig基因表达的启动子,并根据控制其活性的σ因子对这些启动子进行分类。为了分析控制大型调控子(其中还包含在指数生长和非应激条件下表达的基因)的主要σ因子的调控,我们使用了在基本培养基中快速生长的红平红球菌CCM2595培养物。通过初级5'端特异性RNA测序检测了sigA、sigB、sigD、sigE、sigG、sigH、sigJ和sigK基因的转录起始位点(TSS)。位于检测到的TSS上游的启动子由其-35和-10元件定义,这些元件与相关物种谷氨酸棒杆菌和结核分枝杆菌中的这些序列相同或非常相似。通过两种独立技术(体内和体外)证明了不同σ因子对启动子活性的调控。发现所有编码具有胞外功能(ECF)的σ因子的分析sig基因也由额外的管家启动子驱动。基于sig基因启动子的分类,设计了红平红球菌基本σ转录调控网络的模型。