Regulated Intramembrane Proteolysis in Mycobacterium Tuberculosis

描述（由申请人提供）：结核分枝杆菌感染仍然是严重的全球健康危机，需要新的治疗药物和预防策略。鉴于这些要求，发现这种生物体的毒力所需的生物过程对于确定潜在的药物靶点至关重要。这些蛋白中有大量与细胞包膜的发生和维持有关。我们实验室之前的工作表明，Rip1 （Rv2869c）膜内蛋白酶是小鼠生长和持续所必需的，也是维持分枝杆菌细胞膜的必要条件。Rip1是膜内金属蛋白酶（S2P）家族的一员，在其跨膜结构域内切割底物。这些蛋白水解级联反应涉及通过下游转录网络在细胞间传导环境刺激。初步数据表明，Rip1蛋白酶活性受铁限制而激活，并控制多种下游途径。该应用程序描述了一种逻辑方法，其中传统遗传学和生化技术的结合允许鉴定下游转录靶标对小鼠中Rip1依赖性衰减的相对贡献。此外，我们将确定Rip1依赖性低铁信号的分子机制。

DESCRIPTION (provided by applicant): M. tuberculosis infection remains a severe global health crisis that demands new therapeutic agents and preventative strategies. In light of these requirements, the discovery of biological processes required for the virulence of this organism have been paramount for the identification of potential drug targets. A large number of these proteins have been implicated in cell envelope biogenesis and maintenance. Previous work in our laboratory has shown that the Rip1 (Rv2869c) intramembrane protease is required for growth and persistence in mice and is necessary for maintenance of the cell envelope of mycobacteria. Rip1 is a member of the Site two protease (S2P) family of intramembrane metalloproteases which cleave substrates within their transmembrane domains. These proteolytic cascades have been implicated in the transduction of environmental stimuli between cellular compartments via downstream transcriptional networks. Preliminary data show that Rip1 protease activity is activated by iron limitation and controls multiple downstream pathways. This application describes a logical approach in which a combination of conventional genetics and biochemical techniques allow for the identification of the relative contribution of the downstream transcriptional targets to Rip1 dependent attenuation in the mouse. Furthermore, we will determine the molecular mechanism responsible for Rip1 dependent low iron signaling.