Vohradsky Jiri, Schwarz Marek, Ramaniuk Olga, Ruiz-Larrabeiti Olatz, Vaňková Hausnerová Viola, Šanderová Hana, Krásný Libor
Laboratory of Bioinformatics, Institute of Microbiology of the Czech Academy of Sciences, Vídeňská 1083, 14220 Prague, Czech Republic.
Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Vídeňská 1083, 14220 Prague, Czech Republic.
Microorganisms. 2021 Jan 5;9(1):112. doi: 10.3390/microorganisms9010112.
The exponential increase in the number of conducted studies combined with the development of sequencing methods have led to an enormous accumulation of partially processed experimental data in the past two decades. Here, we present an approach using literature-mined data complemented with gene expression kinetic modeling and promoter sequence analysis. This approach allowed us to identify the regulon of sigma factor SigB of RNA polymerase (RNAP) specifically expressed during germination and outgrowth. SigB is critical for the cell's response to general stress but is also expressed during spore germination and outgrowth, and this specific regulon is not known. This approach allowed us to (i) define a subset of the known SigB regulon controlled by SigB specifically during spore germination and outgrowth, (ii) identify the influence of the promoter sequence binding motif organization on the expression of the SigB-regulated genes, and (iii) suggest additional sigma factors co-controlling other SigB-dependent genes. Experiments then validated promoter sequence characteristics necessary for direct RNAP-SigB binding. In summary, this work documents the potential of computational approaches to unravel new information even for a well-studied system; moreover, the study specifically identifies the subset of the SigB regulon, which is activated during germination and outgrowth.
在过去二十年中,所开展研究数量的指数级增长以及测序方法的发展,导致了大量部分处理的实验数据的积累。在此,我们提出一种方法,该方法利用文献挖掘数据,并辅以基因表达动力学建模和启动子序列分析。这种方法使我们能够鉴定出在芽孢萌发和生长过程中特异性表达的RNA聚合酶(RNAP)的σ因子SigB的调控子。SigB对细胞应对一般应激至关重要,但在孢子萌发和生长过程中也有表达,而这个特定的调控子尚不清楚。这种方法使我们能够:(i)定义在孢子萌发和生长过程中由SigB特异性控制的已知SigB调控子的一个子集;(ii)确定启动子序列结合基序组织对SigB调控基因表达的影响;(iii)提出共同控制其他SigB依赖性基因的额外σ因子。随后的实验验证了直接RNAP-SigB结合所需的启动子序列特征。总之,这项工作证明了计算方法即使对于一个研究充分的系统也有挖掘新信息的潜力;此外,该研究具体鉴定了在萌发和生长过程中被激活的SigB调控子的子集。