Department of Microbiology and Molecular Genetics, IMRIC, The Hebrew University of Jerusalem, Faculty of Medicine, Jerusalem, Israel.
Department of Microbiology and Molecular Genetics, IMRIC, The Hebrew University of Jerusalem, Faculty of Medicine, Jerusalem, Israel
J Bacteriol. 2014 Aug;196(15):2798-806. doi: 10.1128/JB.00069-14. Epub 2014 May 16.
Enteropathogenic Escherichia coli (EPEC) is a major cause of food poisoning, leading to significant morbidity and mortality. EPEC virulence is dependent on a type III secretion system (T3SS), a molecular syringe employed by EPEC to inject effector proteins into host cells. The injected effector proteins subvert host cellular functions to the benefit of the infecting bacteria. The T3SS and related genes reside in several operons clustered in the locus of enterocyte effacement (LEE). We carried out simultaneous analysis of the expression dynamics of all the LEE promoters and the rate of maturation of the T3SS. The results showed that expression of the LEE1 operon is activated immediately upon shifting the culture to inducing conditions, while expression of other LEE promoters is activated only ∼70 min postinduction. Parallel analysis showed that the T3SS becomes functional around 100 min postinduction. The T3SS core proteins EscS, EscT, EscU, and EscR are predicted to be involved in the first step of T3SS assembly and are therefore included among the LEE1 genes. However, interfering with the temporal regulation of EscS, EscT, EscU, and EscR expression has only a marginal effect on the rate of the T3SS assembly. This study provides a comprehensive description of the transcription dynamics of all the LEE genes and correlates it to that of T3SS biogenesis.
肠致病性大肠杆菌(EPEC)是食物中毒的主要原因,导致发病率和死亡率显著上升。EPEC 的毒力依赖于 III 型分泌系统(T3SS),这是一种 EPEC 用来将效应蛋白注射到宿主细胞中的分子注射器。注入的效应蛋白颠覆了宿主细胞的功能,有利于感染细菌。T3SS 及相关基因位于几个操纵子中,这些操纵子聚集在肠细胞脱落(LEE)基因座中。我们同时分析了所有 LEE 启动子的表达动态和 T3SS 的成熟速度。结果表明,当培养物转移到诱导条件下时,LEE1 操纵子的表达立即被激活,而其他 LEE 启动子的表达仅在诱导后约 70 分钟被激活。平行分析表明,T3SS 在诱导后约 100 分钟时变得功能化。T3SS 核心蛋白 EscS、EscT、EscU 和 EscR 预计参与 T3SS 组装的第一步,因此被包含在 LEE1 基因中。然而,干扰 EscS、EscT、EscU 和 EscR 表达的时间调控对 T3SS 组装速度只有轻微的影响。这项研究全面描述了所有 LEE 基因的转录动态,并将其与 T3SS 生物发生相关联。