Hahn J, Luttinger A, Dubnau D
Public Health Research Institute, New York, New York 10016, USA.
Mol Microbiol. 1996 Aug;21(4):763-75. doi: 10.1046/j.1365-2958.1996.371407.x.
Competence in Bacillus subtilis is expressed post-exponentially in response to signals which are interpreted by a complex network of regulatory proteins. This network culminates in the transcriptional activation of a set of late-competence proteins that mediate DNA binding and uptake during transformation. ComK, a protein that binds to competence promoters and appears to activate their transcription, is itself synthesized in response to the signal-transduction network. ComK is known to be required for the transcription of its own gene. We have placed comK under control of the xylose-inducible PxylA promoter and used this construct to show that ComK synthesis is sufficient as well as necessary to induce competence. We have also confirmed that the Mec proteins act post-transcriptionally to inactivate ComK, probably by protein-protein interaction. We have further demonstrated that ComS is required to generate an upstream signal that causes reversal of Mec-induced inactivation of ComK. In addition to ComK itself, DegU, AbrB, and SinR are required for comK transcription; mutations in their genes are bypassed by PxylA-comK induction, and therefore their products appear not to act via the Mec proteins. Overproduction of ComK, in a loss-of-function mec mutant, is also known to bypass the need for DegU, SinR and AbrB. We propose that these proteins enhance the activity of ComK as a positive autoregulatory transcription factor, acting as coactivator proteins when ComK is present at low concentrations. Finally, we demonstrate that when ComK is synthesized from the PxylA promoter and mecA is inactivated by mutation, no additional growth-stage-regulated control of competence can be detected.
枯草芽孢杆菌的感受态是在指数生长期后表达的,以响应由复杂的调控蛋白网络所解读的信号。这个网络最终导致一组晚期感受态蛋白的转录激活,这些蛋白在转化过程中介导DNA结合和摄取。ComK是一种与感受态启动子结合并似乎激活其转录的蛋白质,它本身是响应信号转导网络而合成的。已知ComK是其自身基因转录所必需的。我们将comK置于木糖诱导型PxylA启动子的控制之下,并利用这个构建体表明ComK的合成对于诱导感受态既是充分的也是必要的。我们还证实,Mec蛋白在转录后发挥作用使ComK失活,可能是通过蛋白质-蛋白质相互作用。我们进一步证明,ComS是产生上游信号所必需的,该信号导致Mec诱导的ComK失活的逆转。除了ComK本身之外,DegU、AbrB和SinR对于comK转录也是必需的;它们基因中的突变可被PxylA-comK诱导所绕过,因此它们的产物似乎不是通过Mec蛋白发挥作用。在功能丧失的mec突变体中ComK的过量产生也已知可绕过对DegU、SinR和AbrB的需求。我们提出,这些蛋白质增强ComK作为一种正自调控转录因子的活性,当ComK以低浓度存在时作为共激活蛋白发挥作用。最后,我们证明,当ComK从PxylA启动子合成且mecA因突变而失活时,无法检测到对感受态的额外生长阶段调控。