Investigating the function of ZU5 domain-containing proteins as amplifiers of caspase activation

项目摘要和摘要 Caspase-1是一种半胱氨酸蛋白酶，催化细胞因子的成熟，在先天的 以及对致病刺激的适应性免疫反应。Caspase-1的错误调控与多种 自身免疫性疾病和癌症。同样，caspase-2是一种半胱氨酸蛋白酶，对调节 细胞对引起DNA损伤的压力的反应，如化疗。Caspase-1和caspase- 2被结构相似的传感器蛋白激活，这些传感器蛋白感知细胞内的扰动并安装 适当的反应，但感应器如何被激活并随后被激活的分子机制 它们各自的蛋白水解酶，还没有被很好地理解。一系列生殖系编码的模式识别受体 感觉病原体的保守特征，并组装成称为炎症体的多蛋白复合体， 招募并激活caspase-1。Caspase-1激活的共识模型是炎性小体 首先激活，然后它们又激活caspase-1。我们的初步数据表明，caspase-1在 炎性小体激活，进而激活更多的caspase-1，然而，其分子机制是 未知。K99指导阶段的目标是确定caspase-1在炎症体中所起的作用。 激活。具体地说，我们将确定ZU5结构域的激活机制 炎性小体、CARD8和NLRP1。在独立的R00阶段，然后我们将应用来自 指导阶段，以确定另一个包含ZU5结构域的传感器的激活机制， 激活caspase-2以响应基因毒性应激，PIDD。我们的中心假设是ZU5结构域- 含有感受器蛋白的蛋白以类似的方式被激活，它们激活的蛋白水解酶参与其中。 传感器激活，进而激活更多的蛋白水解酶。为了实现这些目标，我精心组装了 一个高度互补的顾问团队，拥有所需的科学和指导技能，指导我的道路 研究独立性。这项工作的完成将加深我们对上睑下垂和细胞凋亡的理解 监管，并有可能推动各种人类疾病的治疗开发工作。

PROJECT SUMMARY AND ABSTRACT Caspase-1 is a cysteine protease that catalyzes the maturation of cytokines and plays critical roles in the innate and adaptive immune response to pathogenic stimuli. Misregulation of caspase-1 is associated with various autoimmune diseases and cancer. Similarly, caspase-2 is a cysteine protease that is important for regulating the cellular response to stresses that cause DNA damage, such as chemotherapy. Both caspase-1 and caspase- 2 are activated by structurally similar sensor proteins that sense intracellular perturbations and mount appropriate responses, but the molecular mechanism of how the sensors are activated and then in turn activate their respective proteases, is not well understood. A series of germline-encoded pattern recognition receptors sense conserved features of pathogens, and assemble into multiprotein complexes called inflammasomes, which recruit and activate caspase-1. The consensus model for caspase-1 activation is that inflammasomes are first activated then they in turn activate caspase-1. Our preliminary data suggests that caspase-1 plays a role in inflammasome activation, which in turn activates more caspase-1, however, the molecular mechanism is unknown. The goal during the K99 mentored phase, is to determine the role caspase-1 plays in inflammasome activation. Specifically, we will determine the activation mechanism of the ZU5 domain-containing inflammasomes, CARD8 and NLRP1. During the independent R00 phase, we will then apply the training from the mentored phase to determine the activation mechanism of another ZU5 domain-containing sensor that activates caspase-2 in response to genotoxic stress, PIDD. Our central hypothesis is that the ZU5 domain- containing sensor proteins are activated in a similar manner, in which the proteases they activate participate in sensor activation, which in turn activates more protease. To accomplish these goals, I have carefully assembled a highly complementary advisory team with the scientific and mentoring skills needed to guide my path to research independence. The completion of this work will further our understanding of pyroptosis and apoptosis regulation and could potentially advance therapeutic development efforts for a variety of human diseases.