Department of Cell Biology, Microbiology and Molecular Biology, University of South Florida, 4202 E. Fowler Ave, Tampa, FL, 33620, USA.
Department of Biological Sciences, Ohio University, Athens, OH, 45701, USA.
FEMS Microbiol Lett. 2018 Jun 1;365(12). doi: 10.1093/femsle/fny108.
Staphylococcus aureus possess three alternative σ factors, including a lone extracytoplasmic function σ factor, σS. Our group previously identified and characterized this element, mapping three sigS promoters, demonstrating its inducibility during stress and virulence inducing conditions and demonstrating a role for this factor in disease causation. In the present study, we identify a fourth promoter of the sigS operon, termed P4, located in a unique position internal to the sigS coding region. Transcriptional profiling revealed that expression from P4 is dominant to the three upstream promoters, particularly upon exposure to chemical stressors that elicit DNA damage and disrupt cell wall stability; each of which have previously been shown to stimulate sigS expression. Importantly, expression of this fourth promoter, followed by at least one or more of the upstream promoters, is induced during growth in serum and upon phagocytosis by RAW 264.7 murine macrophage-like cells. Finally, we demonstrate that a downstream gene, SACOL1829, bears a large 3΄ UTR that spans the sigS-SACOL1828 coding region, and may serve to compete with the P4 transcript to inhibit σS production. Collectively, these findings reveal a unique operon architecture for the sigS locus that indicates the potential for novel regulatory mechanisms governing its expression.
金黄色葡萄球菌拥有三种替代的 σ 因子,包括一个独特的细胞外功能 σ 因子 σS。我们的研究小组之前已经鉴定并表征了这个元件,定位了三个 sigS 启动子,证明了它在应激和毒力诱导条件下的诱导性,并证明了该因子在疾病发病机制中的作用。在本研究中,我们鉴定了 sigS 操纵子的第四个启动子 P4,它位于 sigS 编码区内部的独特位置。转录谱分析显示,P4 的表达相对于三个上游启动子占主导地位,特别是在暴露于化学应激剂引起 DNA 损伤和破坏细胞壁稳定性时;这些应激剂之前都被证明可以刺激 sigS 的表达。重要的是,在血清中生长和被 RAW 264.7 鼠巨噬样细胞吞噬时,至少一个或多个上游启动子的表达紧随其后,诱导了这个第四个启动子的表达。最后,我们证明下游基因 SACOL1829 具有一个跨越 sigS-SACOL1828 编码区的大 3'UTR,可能通过与 P4 转录本竞争来抑制 σS 的产生。总的来说,这些发现揭示了 sigS 基因座独特的操纵子结构,表明其表达可能存在新的调控机制。