The Ubiquitin-Proteasome System in Metazoan Embryogenesis

描述（由申请人提供）：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许多理解都围绕着基因调控网络，这些基因调控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）的蛋白水解已经成为在发育的特定阶段去除特定信号分子和转录调节因子以确保发育事件的适当进展的重要调节层。利用C.作为一个模型，我们将采取一种新的多学科的方法，系统和快速阐明UPS在后生动物发育中的体内功能。我们已经确定了约50个保守的，胚胎致死基因所需的UPS底物特异性。此外，我们还开发了新技术，可以在胚胎发生的每一分钟自动跟踪每个细胞，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系统地分析每个细胞行为的方法。使用这些技术，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，我们将采取系统生物学的方法来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的基础上表型相似性，并整合这个发展网络与现有的大型人类基因相互作用网络的基础上序列的同源性。在这样做的过程中，我们将建立一个广泛的基因网络，将UPS基因及其相互作用物与特定的发育过程及其潜在的调控途径联系起来。因此，该网络将特别有助于指导未来关于UPS如何调节人类发育和疾病的研究。最后，我们将深入研究在C. elegans胚胎，C. elegans的发展揭示了我们的UPS的试点研究。

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.