Mechanisms regulating nuclear envelope structure and stability

项目总结 长期以来，核膜(NE)一直被认为在间期起静态屏障的作用 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破裂会导致暂时或永久性的细胞核区域化丧失。 在细胞分裂和迁移中观察到，并在椎板病的细胞中出现频率增加 和癌症患者。目前的研究表明，东北方向破裂可能会产生重大后果 基因表达、染色质结构、非整倍体和基因组稳定性。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 定义细胞后果。这在一定程度上是因为我们对NE的结构了解有限， 尤其是层次化的网络组织，以及结构变化是如何调节的。我们当前的目标是 研究计划是1)确定NE破裂和修复的机制，2)开发新的工具来 分析NE结构。我们计划通过在癌症中进行全基因组RNAi筛查来实现这些目标 确定去甲肾上腺素稳定性的调节因素，并设计新的图像分析程序和活细胞成像 记者研究了NE的3D动力学和物理性质。我们正处于定义的开始 NE作为一种动态结构。因此，这项研究计划的长期目标是确定 调节去甲肾上腺素动力学的基本原理和确定缺陷的分子机制 Ne稳定性影响疾病。确定调节NE断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，转录调节，细胞对机械的反应 力量、DNA复制和修复，以及关于细胞分化、组织发育、 衰老，以及癌症的发生和发展。

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.