Regulated Intramembrane Proteolysis in Mycobacterium Tuberculosis

描述（由申请人提供）：M。结核病感染仍然是严重的全球健康危机，需要新的治疗剂和预防策略。鉴于这些要求，发现该生物体毒力所需的生物学过程对于鉴定潜在的药物靶点至关重要。大量的这些蛋白质已经涉及细胞包膜的生物发生和维持。我们实验室以前的工作表明，Rip1（Rv2869c）膜内蛋白酶是小鼠生长和持久性所必需的，也是维持分枝杆菌细胞包膜所必需的。Rip1是膜内金属蛋白酶的位点2蛋白酶（S2P）家族的成员，其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区室之间转导环境刺激。初步数据显示，Rip1蛋白酶活性被铁限制激活，并控制多个下游途径。本申请描述了一种逻辑方法，其中常规遗传学和生物化学技术的组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.