Xavier Karina B, Bassler Bonnie L
Department of Molecular Biology, Princeton University, Princeton, NJ 08544-1014, USA.
J Bacteriol. 2005 Jan;187(1):238-48. doi: 10.1128/JB.187.1.238-248.2005.
AI-2 is a quorum-sensing signaling molecule proposed to be involved in interspecies communication. In Escherichia coli and Salmonella enterica serovar Typhimurium, extracellular AI-2 accumulates in exponential phase, but the amount decreases drastically upon entry into stationary phase. In S. enterica serovar Typhimurium, the reduction in activity is due to import and processing of AI-2 by the Lsr transporter. We show that the Lsr transporter is functional in E. coli, and screening for mutants defective in AI-2 internalization revealed lsrK and glpD. Unlike the wild type, lsrK and glpD mutants do not activate transcription of the lsr operon in response to AI-2. lsrK encodes the AI-2 kinase, and the lsrK mutant fails to activate lsr expression because it cannot produce phospho-AI-2, which is the lsr operon inducer. glpD encodes the glycerol-3-phosphate (G3P) dehydrogenase, which is involved in glycerol and G3P metabolism. G3P accumulates in the glpD mutant and represses lsr transcription by preventing cyclic AMP (cAMP)-catabolite activator protein (CAP)-dependent activation. Dihydroxyacetone phosphate (DHAP) also accumulates in the glpD mutant, and DHAP represses lsr transcription by a cAMP-CAP-independent mechanism involving LsrR, the lsr operon repressor. The requirement for cAMP-CAP in lsr activation explains why AI-2 persists in culture fluids of bacteria grown in media containing sugars that cause catabolite repression. These findings show that, depending on the prevailing growth conditions, the amount of time that the AI-2 signal is present and, in turn, the time that a given community of bacteria remains exposed to this signal can vary greatly.
AI-2是一种群体感应信号分子,被认为参与种间通讯。在大肠杆菌和肠炎沙门氏菌血清型鼠伤寒沙门氏菌中,细胞外AI-2在指数生长期积累,但进入稳定期后数量急剧下降。在鼠伤寒沙门氏菌中,活性降低是由于Lsr转运蛋白对AI-2的导入和加工。我们发现Lsr转运蛋白在大肠杆菌中具有功能,通过筛选AI-2内化缺陷的突变体发现了lsrK和glpD。与野生型不同,lsrK和glpD突变体在响应AI-2时不会激活lsr操纵子的转录。lsrK编码AI-2激酶,lsrK突变体无法激活lsr表达,因为它不能产生磷酸化AI-2,而磷酸化AI-2是lsr操纵子的诱导剂。glpD编码甘油-3-磷酸(G3P)脱氢酶,其参与甘油和G3P代谢。G3P在glpD突变体中积累,并通过阻止环磷酸腺苷(cAMP)-分解代谢物激活蛋白(CAP)依赖性激活来抑制lsr转录。磷酸二羟丙酮(DHAP)也在glpD突变体中积累,并且DHAP通过一种不依赖cAMP-CAP的机制抑制lsr转录,该机制涉及lsr操纵子阻遏物LsrR。lsr激活对cAMP-CAP的需求解释了为什么AI-2会在含有导致分解代谢物阻遏的糖类的培养基中生长的细菌培养液中持续存在。这些发现表明,根据当时的生长条件,AI-2信号存在的时间,进而给定细菌群体暴露于该信号的时间可能会有很大差异。