Mechanisms regulating nuclear envelope structure and stability

项目摘要 长期以来，核被膜（NE）被认为是间期的静态屏障， 调节胞质蛋白和染色质之间的相互作用。然而，最近的工作挑战了这一点。 假设NE破裂，导致短暂或永久性的核区室化丧失， 在分裂和迁移的细胞中观察到，在核纤层蛋白病的细胞中发生频率增加 和癌症患者。目前的研究表明，NE破裂可能会产生重大后果 基因表达、染色质结构、非整倍性和基因组稳定性。然而，我们几乎没有 关于驱动NE破裂和修复的机制的信息，因此，我们无法严格地 定义细胞的后果。这在一定程度上是由于我们对NE结构的了解有限， 特别是核纤层蛋白网络组织，以及结构变化是如何调节的。我们当前的目标 研究计划是1）确定NE断裂和修复的机制，2）开发新的工具， 分析NE结构。我们计划通过在癌症中进行全基因组RNAi筛选来实现这些目标 细胞，以确定NE稳定性的调节剂，并设计新的图像分析程序和活细胞成像 研究NE的3D动力学和物理特性。我们刚刚开始定义 作为一个动态的结构。因此，本研究计划的长期目标是确定 调节NE动力学的基本原理，并确定了 NE稳定性影响疾病。确定调节NE破裂和修复的蛋白质和途径将 重新定义我们对NE结构的理解，并对相关领域产生广泛影响，包括 染色体组织和染色质结构，转录调控，细胞对机械 力，DNA复制和修复，以及关于细胞分化，组织发育， 衰老和癌症的发生和发展。

PROJECT SUMMARY The nuclear envelope (NE) has long been thought to function as a static barrier during interphase that regulates interactions between cytosolic proteins and the chromatin. However, recent work challenges this assumption. NE rupture, which results in the transient or permanent loss of nucleus compartmentalization, has been observed in dividing and migrating cells and occurs with increased frequency in cells from laminopathy and cancer patients. Current research has demonstrated that NE rupture can have significant consequences for gene expression, chromatin structure, aneuploidy, and genome stability. However, we have almost no information about the mechanisms driving NE rupture and repair and, thus, we have been unable to rigorously define the cellular consequences. This is due, in part, to our limited understanding of the structure of the NE, particularly lamin network organization, and how structural changes are regulated. The goals of our current research program are 1) to identify the mechanisms of NE rupture and repair, and 2) to develop new tools to analyze NE structure. We plan to achieve these goals by performing a genome-wide RNAi screen in cancer cells to identify regulators of NE stability, and designing new image analysis programs and live-cell imaging reporters to study the 3D dynamics and physical properties of the NE. We are at the beginning of defining the NE as a dynamic structure. Thus, the long-term goals of this research program are to identify the fundamental principles regulating NE dynamics and determine the molecular mechanisms by which defects in NE stability impact disease. Identifying the proteins and pathways that regulate NE rupture and repair will redefine our understanding of NE structure and have broad impacts on associated fields, including chromosome organization and chromatin structure, transcription regulation, cellular responses to mechanical force, and DNA replication and repair, and on larger questions about cell differentiation, tissue development, aging, and cancer initiation and progression.