The Ubiquitin-Proteasome System in Metazoan Embryogenesis

描述(申请人提供)：后生动物胚胎发生中的泛素-蛋白酶体系统我们对发育的遗传调控的大部分理解都集中在基因调控网络上，该网络驱动具有时间和空间特异性的不同转录级联。通过泛素-蛋白酶体系统(UPS)的蛋白分解已成为重要的调节层，在发育的特定阶段移除特定的信号分子和转录调节因子，以确保发育事件的正常进行。以线虫胚胎发生为模型，我们将采用一种新的多学科方法来系统和快速地阐明UPS在后生动物发育中的体内功能。我们已经确定了大约50个保守的胚胎致死基因，这些基因是UPS底物特异性所必需的。此外，我们还开发了在胚胎发生过程中每时每刻自动跟踪每个细胞的新技术，并将进一步开发基于细胞分裂、命运标记表达和细胞运动的定量测量来系统分析每个细胞行为的方法。利用这些技术，我们的遗传和表型分析将定义这些~50个保守基因在单细胞分辨率下调节的特定发育背景和过程。然后，我们将采用系统生物学的方法，基于表型相似性，构建这些基因与关键发育调控因子和途径的预测相互作用网络，并将该发育网络与基于序列正交学的现有大型人类基因相互作用网络整合。在这样做的过程中，我们将建立一个广泛的基因网络，将UPS基因及其相互作用者与特定的发育过程及其潜在的调控途径联系起来。因此，这个网络在指导未来关于UPS如何监管人类发展和疾病的研究中将特别有用。最后，我们将深入研究UPS如何在线虫早期胚胎中控制自我更新和分化之间的平衡，这是线虫发育的一个新颖和意想不到的方面，在我们对UPS的初步研究中揭示了这一点。 公共卫生相关性：泛素蛋白酶体系统是细胞中的一个复杂的调控网络。使用系统生物学战略和涉及活显微镜、遗传学和计算科学的多学科方法，我们将把这些关键调控因子与发育过程和人类病理学联系起来，例如癌症中的过度增殖。

DESCRIPTION (provided by applicant): The Ubiquitin-Proteasome System in Metazoan Embryogenesis Much of our understanding about the genetic regulation of development centers around gene regulatory networks that drive different transcriptional cascades with temporal and spatial specificity. Proteolysis through the ubiquitin-proteasome system (UPS) has emerged as an essential layer of regulation to remove specific signaling molecules and transcriptional regulators at specific stages of development to ensure proper progression of developmental events. Using C. elegans embryogenesis as a model, we will take a novel multidisciplinary approach to systematically and rapidly elucidate the in vivo functions of the UPS in Metazoan development. We have identified ~50 conserved, embryonic lethal genes that are required for UPS substrate specificity. In addition, we have developed novel technologies to automatically track every cell at every minute through embryogenesis, and will further develop methods to systematically assay the behaviors of every cell based on quantitative measurements of cell division, fate marker expression and cell movement. Using these technologies, our genetic and phenotypic analysis will define the specific developmental contexts and processes that these ~50 conserved genes regulate at single-cell resolution. We will then take a systems biology approach to construct a predicted interaction network of these genes with key developmental regulators and pathways based on phenotypic similarity, and integrate this developmental network with the large existing human gene interaction network based on sequence orthology. In doing so, we will build an extensive gene network that connects UPS genes and their interactors to specific developmental processes and their underlying regulatory pathways. This network will therefore be particularly useful in guiding future studies of how the UPS regulates human development and disease. Finally, we will conduct an in depth study on how the UPS controls the balance between self renewal and differentiation in the early C. elegans embryo, a novel and unexpected aspect of C. elegans development revealed in our pilot studies of the UPS. PUBLIC HEALTH RELEVANCE: The Ubiquitin Proteasome System is a complex regulatory network in cells. Using a Systems Biology strategy and a multidisciplinary approach involving live microscopy, genetics and computational sciences, we will link these key regulators to developmental processes and human pathology, such as over proliferation in cancer.