Muller Cécile, Le Breton Yoann, Morin Thierry, Benachour Abdellah, Auffray Yanick, Rincé Alain
USC INRA 2017 Microbiologie de l'Environnement, EA 956, IRBA, Université de Caen, 14032 CAEN cedex, France.
J Bacteriol. 2006 Apr;188(7):2636-45. doi: 10.1128/JB.188.7.2636-2645.2006.
The Enterococcus faecalis two-component signal transduction system CroRS, also referred as the RR-HK05 pair, is required for intrinsic beta-lactam resistance (Y. R. Comenge, R. Quintiliani, Jr., L. Li, L. Dubost, J. P. Brouard, J. E. Hugonnet, and M. Arthur, J. Bacteriol. 185:7184-7192, 2003) and is also suspected to be involved in the expression of salB (previously referred to as sagA), a gene important for resistance to environmental stress and cell morphology (Y. Le Breton, G. Boël, A. Benachour, H. Prévost, Y. Auffray, and A. Rincé, Environ. Microbiol. 5:329-337, 2003). In this report, we provide genetic and biochemical evidence that salB encodes a secreted protein that is expressed from a monocistronic stress-inducible operon. Consistent with CroR being a direct transcriptional activator of the salB expression, CroR was found to bind to the salB promoter region in electrophoretic mobility shift assays. Interestingly, we provide evidence that SalB does not play a role in the intrinsic beta-lactam resistance associated with CroRS. We also show that the CroRS system is able to regulate its own expression. The sequence of the CroRS binding site in the salB and croR promoter regions was determined using DNase I footprinting assays.
粪肠球菌双组分信号转导系统CroRS,也被称为RR-HK05对,是固有β-内酰胺抗性所必需的(Y. R. Comenge、R. Quintiliani, Jr.、L. Li、L. Dubost、J. P. Brouard、J. E. Hugonnet和M. Arthur,《细菌学杂志》185:7184 - 7192,2003年),并且还被怀疑参与salB(以前称为sagA)的表达,salB是一个对抵抗环境压力和细胞形态很重要的基因(Y. Le Breton、G. Boël、A. Benachour、H. Prévost、Y. Auffray和A. Rincé,《环境微生物学》5:329 - 337,2003年)。在本报告中,我们提供了遗传和生化证据,表明salB编码一种从单顺反子应激诱导操纵子表达的分泌蛋白。与CroR是salB表达的直接转录激活因子一致,在电泳迁移率变动分析中发现CroR与salB启动子区域结合。有趣的是,我们提供的证据表明SalB在与CroRS相关的固有β-内酰胺抗性中不起作用。我们还表明CroRS系统能够调节其自身的表达。使用DNase I足迹分析确定了salB和croR启动子区域中CroRS结合位点的序列。