Identification of rovA Regulators in Y.Enterocolitica

描述(申请人提供)：耶尔森氏菌属包含三种致病物种。小肠结肠炎耶尔森氏菌和假结核耶尔森氏菌是引起胃肠炎的肠道病原体，鼠疫杆菌是鼠疫的病原体。耶尔森氏菌RoVa蛋白是细菌全局调节因子MarR/SlyA家族的成员，最初被确定为inv的正调节因子，inv是编码inasin的基因，invasin是感染初期参与的一种关键毒力因子。RoVa是完全毒力所必需的，并且RoVa突变体比inv突变体更弱，这表明RoVa可能调节毒力所需的其他基因。RoVa对多种毒力因子的调控为介导毒力提供了一个中央控制点，也为控制几个基因的表达提供了一个调控检查点。因此，诱导或抑制RoVa表达的环境因素将反过来同时调节几个基因，从而允许协调建立感染所需的各种因素的表达。在这里，我建议确定RoVa在感染过程中的表达模式，并确定调节RoVa表达的环境因素。此外，RoVa调节子将通过转座子突变和启动子结合分析来确定。最后，我建议确定RoVa调节器是否与RoVa启动子相互作用，以及RoVa调节器在毒力中的作用。这些目标的完成将极大地提高我们对小肠结肠炎耶尔森氏菌中一个重要毒力因子的调控的了解，并可能识别耶尔森氏菌中新的毒力调控途径。

DESCRIPTION (provided by applicant): The genus Yersinia contains three pathogenic species. Y. enterocolitica and Y. pseudotuberculosis are enteric pathogens that cause gastroenteritis, and Y. pestis is the causative agent of plague. The Yersinia RovA protein is a member of the MarR/SlyA family of bacterial global regulators and was first identified as a positive regulator of inv, the gene that encodes invasin, a critical virulence factor involved in the initial stages of infection. RovA is necessary for full virulence and a rovA mutant is more attenuated than an inv mutant, which indicates that RovA may regulate other genes required for virulence. The regulation of multiple virulence factors by RovA provides a central control point to mediate virulence and a regulatory checkpoint to control expression of several genes. Therefore, environmental factors that induce or repress expression of rovA will in turn regulate several genes at once, allowing for the coordination of expression of factors necessary to establish infection. Here I propose to identify the expression pattern of rovA during infection and to determine the environmental factors that regulate rovA expression. Furthermore, rovA regulators will be identified using transposon mutagenesis and promoter binding assays. Finally, I propose to determine whether rovA regulators interact with the rovA promoter and the role of rovA regulators in virulence. Completion of these aims will greatly enhance our knowledge of the regulation of an important virulence factor in Y. enterocolitica and may identify novel virulence regulatory pathways in Yersinia.