Mandic-Mulec I, Doukhan L, Smith I
Public Health Research Institute, New York, New York 10016, USA.
J Bacteriol. 1995 Aug;177(16):4619-27. doi: 10.1128/jb.177.16.4619-4627.1995.
SinR is a pleiotropic DNA binding protein that is essential for the late-growth processes of competence and motility in Bacillus subtilis and is also a repressor of others, e.g., sporulation and subtilisin synthesis. In this report, we show that SinR, in addition to being an inhibitor of sporulation stage II gene expression, is a repressor of the key early sporulation gene spo0A. The sporulation-specific rise in spo0A expression at time zero is absent in a SinR-overproducing strain and is much higher than normal in strains with a disrupted sinR gene. This effect is direct, since SinR binds specifically to spo0A in vitro, in a region overlapping the -10 region of the sporulation-specific Ps promoter that is recognized by E-sigma H polymerase. Methyl interference and site-directed mutagenesis studies have identified guanine residues that are important for SinR recognition of this DNA sequence. Finally, we present evidence that SinR controls sporulation through several independent genes, i.e., sp0A, spoIIA, and possibly spoIIG and spoIIE.
SinR是一种多效性DNA结合蛋白,对枯草芽孢杆菌感受态和运动性的后期生长过程至关重要,同时也是其他过程(如芽孢形成和枯草杆菌蛋白酶合成)的阻遏物。在本报告中,我们表明,SinR除了是芽孢形成II期基因表达的抑制剂外,还是关键的早期芽孢形成基因spo0A的阻遏物。在SinR过量表达的菌株中,芽孢形成特异性的spo0A表达在时间为零时的上升不存在,而在sinR基因被破坏的菌株中,其表达比正常情况高得多。这种效应是直接的,因为SinR在体外特异性结合spo0A,结合区域与芽孢形成特异性Ps启动子的-10区域重叠,该启动子可被E-σH聚合酶识别。甲基干扰和定点诱变研究已经确定了对SinR识别该DNA序列很重要的鸟嘌呤残基。最后,我们提供证据表明,SinR通过几个独立的基因(即sp0A、spoIIA,可能还有spoIIG和spoIIE)控制芽孢形成。