Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA.
Appl Environ Microbiol. 2010 Aug;76(15):4996-5004. doi: 10.1128/AEM.00886-10. Epub 2010 Jun 11.
The type 3 secretion system (T3SS) genes of Vibrio harveyi are activated at low cell density and repressed at high cell density by quorum sensing (QS). Repression requires LuxR, the master transcriptional regulator of QS-controlled genes. Here, we determine the mechanism underlying the LuxR repression of the T3SS system. Using a fluorescence-based cell sorting approach, we isolated V. harveyi mutants that are unable to express T3SS genes at low cell density and identified two mutations in the V. harveyi exsBA operon. While LuxR directly represses the expression of exsBA, complementation and epistasis analyses reveal that it is the repression of exsA expression, but not exsB expression, that is responsible for the QS-mediated repression of T3SS genes at high cell density. The present work further defines the genes in the V. harveyi QS regulon and elucidates a mechanism demonstrating how multiple regulators can be linked in series to direct the expression of QS target genes specifically at low or high cell density.
Harveyi 型 3 分泌系统 (T3SS) 基因受群体感应 (QS) 调控,在低细胞密度时被激活,在高细胞密度时被抑制。抑制作用需要 LuxR,这是 QS 控制基因的主要转录调节剂。在这里,我们确定了 LuxR 对 T3SS 系统的抑制机制。我们使用基于荧光的细胞分选方法,分离出不能在低细胞密度下表达 T3SS 基因的 Harveyi 突变体,并在 Harveyi exsBA 操纵子中鉴定出两个突变。虽然 LuxR 直接抑制 exsBA 的表达,但互补和上位性分析表明,是 exsA 表达的抑制,而不是 exsB 表达的抑制,导致高细胞密度时 QS 介导的 T3SS 基因抑制。本工作进一步确定了 Harveyi QS 调控基因,并阐明了一种机制,表明多个调节剂如何串联起来,特异性地在低或高细胞密度下指导 QS 靶基因的表达。