Department of Biology, University of Bologna, Via Selmi 3, 40126 Bologna, Italy.
J Bacteriol. 2011 Oct;193(20):5629-36. doi: 10.1128/JB.05295-11. Epub 2011 Aug 12.
The ability of pathogens to cope with disparate environmental stresses is a crucial feature for bacterial survival and for the establishment of a successful infection and colonization of the host; in this respect, chaperones and heat shock proteins (HSPs) play a fundamental role in host-pathogen interactions. In Helicobacter pylori, the expression of the major HSPs is tightly regulated through dedicated transcriptional repressors (named HspR and HrcA), as well as via a GroESL-dependent posttranscriptional feedback control acting positively on the DNA binding affinity of the HrcA regulator itself. In the present work we show that the CbpA chaperone also participates in the posttranscriptional feedback control of the H. pylori heat shock regulatory network. Our experiments suggest that CbpA specifically modulates HspR in vitro binding to DNA without affecting HrcA regulator activity. In particular, CbpA directly interacts with HspR, preventing the repressor from binding to its target operators. This interaction takes place only when HspR is not bound to DNA since CbpA is unable to affect HspR once the repressor is bound to its operator site. Accordingly, in vivo overexpression of CbpA compromises the response kinetics of the regulatory circuit, inducing a failure to restore HspR-dependent transcriptional repression after heat shock. The data presented in this work support a model in which CbpA acts as an important modulator of HspR regulation by fine-tuning the shutoff response of the regulatory circuit that governs HSP expression in H. pylori.
病原体应对不同环境压力的能力是细菌生存以及成功感染和定植宿主的关键特征;在这方面,伴侣蛋白和热休克蛋白(HSPs)在宿主-病原体相互作用中起着至关重要的作用。在幽门螺杆菌中,主要 HSPs 的表达通过专门的转录抑制剂(称为 HspR 和 HrcA)以及通过 GroESL 依赖性的转录后反馈控制进行严格调节,该反馈控制对 HrcA 调节剂本身的 DNA 结合亲和力具有积极作用。在本工作中,我们表明伴侣蛋白 CbpA 也参与了幽门螺杆菌热休克调控网络的转录后反馈控制。我们的实验表明,CbpA 特异性地调节体外 HspR 与 DNA 的结合,而不影响 HrcA 调节剂的活性。具体来说,CbpA 直接与 HspR 相互作用,阻止抑制剂与其靶标操纵子结合。这种相互作用仅在 HspR 未与 DNA 结合时发生,因为一旦抑制剂与其操纵子结合,CbpA 就无法影响 HspR。因此,体内过表达 CbpA 会破坏调节回路的响应动力学,导致在热休克后无法恢复 HspR 依赖性转录抑制。本工作中提出的数据支持了这样一种模型,即 CbpA 通过微调调节 HSP 表达的调控回路的关闭响应,作为 HspR 调节的重要调节剂。