Luo Ping, Li Haiying, Morrison Donald A
Laboratory for Molecular Biology, Department of Biological Sciences, University of Illinois at Chicago, Chicago, IL 60607, USA.
Mol Microbiol. 2004 Oct;54(1):172-83. doi: 10.1111/j.1365-2958.2004.04254.x.
Regulation of competence for genetic transformation in Streptococcus pneumoniae depends on a quorum-sensing system, genes involved in DNA uptake and recombination and a link between these two gene sets. The alternative sigma factor ComX provides this link. ComE, the response regulator of the quorum-sensing system, is required for expression of ComX and other early genes. However, an unknown ComE-dependent regulator is also required for competence when comX is expressed under control of the raffinose-responsive promoter of the aga operon. The gene comW (SP0018) is required for a high level of competence and is regulated by the quorum-sensing system, but its function is unknown. To explore its role further, comW was cloned into the multicopy plasmid pMSP3535, under the control of a nisin-inducible promoter (P(N)), and transformed into pneumococcal strains containing a raffinose-inducible comX gene (P(R)::comX). Further introduction of a comE deletion blocked the endogenous CSP signal transduction pathway. In the resulting strain, competence was independent of CSP but depended on treatment with both nisin and raffinose, showing that coexpression of comW and comX complemented the comE deficiency. ComX protein accumulation and expression of a late competence gene in the above strain support the conclusion that ComW is a new positive factor involved in competence regulation.
肺炎链球菌中遗传转化能力的调控取决于群体感应系统、参与DNA摄取和重组的基因以及这两组基因之间的联系。替代sigma因子ComX提供了这种联系。群体感应系统的应答调节因子ComE是ComX和其他早期基因表达所必需的。然而,当comX在aga操纵子的棉子糖应答启动子控制下表达时,能力的获得还需要一个未知的ComE依赖性调节因子。基因comW(SP0018)是高水平能力获得所必需的,并且受群体感应系统调控,但其功能尚不清楚。为了进一步探究其作用,将comW克隆到多拷贝质粒pMSP3535中,置于乳链菌肽诱导型启动子(P(N))的控制下,并转化到含有棉子糖诱导型comX基因(P(R)::comX)的肺炎球菌菌株中。进一步引入comE缺失会阻断内源性CSP信号转导途径。在所得菌株中,能力的获得不依赖于CSP,但依赖于乳链菌肽和棉子糖的处理,这表明comW和comX的共表达弥补了comE的缺陷。上述菌株中ComX蛋白的积累和晚期能力基因的表达支持了ComW是参与能力调控的一个新的正调控因子这一结论。