Institute of Molecular Biology, Slovak Academy of Sciences, 845 51 Bratislava, Slovakia.
Int J Mol Sci. 2021 Jul 22;22(15):7849. doi: 10.3390/ijms22157849.
In contrast to , A3(2) contains nine homologues of stress response sigma factor SigB with a major role in differentiation and osmotic stress response. The aim of this study was to further characterize these SigB homologues. We previously established a two-plasmid system to identify promoters recognized by sigma factors and used it to identify promoters recognized by the three SigB homologues, SigF, SigG, and SigH from A3(2). Here, we used this system to identify 14 promoters recognized by SigB. The promoters were verified in vivo in A3(2) under osmotic stress conditions in and operon mutants, indicating some cross-recognition of these promoters by these two SigB homologues. This two-plasmid system was used to examine the recognition of all identified SigB-, SigF-, SigG-, and SigH-dependent promoters with all nine SigB homologues. The results confirmed this cross-recognition. Almost all 24 investigated promoters were recognized by two or more SigB homologues and data suggested some distinguishing groups of promoters recognized by these sigma factors. However, analysis of the promoters did not reveal any specific sequence characteristics for these recognition groups. All promoters showed high similarity in the -35 and -10 regions. Immunoblot analysis revealed the presence of SigB under osmotic stress conditions and SigH during morphological differentiation. Together with the phenotypic analysis of and operon mutants in A3(2), the results suggest a dominant role for SigB in the osmotic stress response and a dual role for SigH in the osmotic stress response and morphological differentiation. These data suggest a complex regulation of the osmotic stress response in relation to morphological differentiation in A3(2).
与 相反,A3(2) 含有九个应激反应σ因子 SigB 的同源物,它们在分化和渗透应激反应中起主要作用。本研究的目的是进一步表征这些 SigB 同源物。我们之前建立了一个双质粒系统来识别 σ 因子识别的启动子,并利用它来识别来自 A3(2) 的三个 SigB 同源物 SigF、SigG 和 SigH 识别的启动子。在这里,我们使用该系统鉴定了 14 个 SigB 识别的启动子。在 和 操纵子突变体的渗透胁迫条件下,在 A3(2) 中体内验证了这些启动子,表明这两个 SigB 同源物对这些启动子存在一些交叉识别。该双质粒系统用于用所有九个 SigB 同源物检查所有鉴定的 SigB、SigF、SigG 和 SigH 依赖性启动子的识别。结果证实了这种交叉识别。几乎所有 24 个研究的启动子都被两个或更多 SigB 同源物识别,并且数据表明这些σ因子识别的一些区分的启动子组。然而,对启动子的分析没有揭示出这些识别组的任何特定序列特征。所有启动子在 -35 和 -10 区域都具有高度相似性。免疫印迹分析显示 SigB 在渗透胁迫条件下存在,SigH 在形态分化过程中存在。结合 A3(2) 中 和 操纵子突变体的表型分析,结果表明 SigB 在渗透应激反应中起主导作用,SigH 在渗透应激反应和形态分化中起双重作用。这些数据表明,A3(2) 中渗透应激反应与形态分化的关系存在复杂的调控。