Chromatin Domain Dynamics

描述（由申请人提供）：细胞核中的DNA与蛋白质络合形成染色质，染色质可变可及性结构域位于细胞核的特定区域。这种包装和组织对于正常的染色体过程（如转录）很重要，已知这种组织的破坏会导致发育缺陷和疾病状态。该建议的总体目标是达到真核生物染色体染色质结构域的结构和组织机制的理解。DNA元素是形成染色质结构域所必需的，这些元素也在细胞核的三维环境中组织结构域。了解DNA元素之间的远程相互作用及其在染色质结构域动力学中的作用是本提案的重点之一。最终目标是了解染色质结合因子和DNA元件如何影响染色质结构，从而影响转录等过程。细胞核中相邻的染色质结构域通常具有拮抗的结构或功能特性，DNA绝缘体元件将这些结构域分开。特定基因的启动子通常在酵母和果蝇中起绝缘子的作用，我们已经确定了这些绝缘子在酵母中起作用的机制的大致轮廓。在这项资助中，我们建议研究细胞分裂时染色质结构域发生的变化，并确定是否有类似的机制在人类细胞中分离染色质结构域。这项研究将涉及分子遗传学和基因组学的使用，使用定量染色质免疫沉淀等技术对染色质进行操作，以及染色体构象捕获和生化分析。了解细胞核是如何组织的应该有助于我们更好地操纵过程，有利于研究。这项研究的结果将有助于理解由染色质结构域错误调控引起的疾病状态的分子基础。研究结果还将使研究人员能够通过开发用于基因治疗的新分子工具来开发治疗疾病状态的新策略。

DESCRIPTION (provided by applicant): The DNA in the nucleus is complexed with proteins to form chromatin and chromatin domains of variable accessibility reside in specific regions of the nucleus. This packaging and organization is important for normal chromosomal processes like transcription and disruption of this organization is known to lead to developmental defects and disease states. The overall objective of this proposal is to reach a mechanistic understanding of the structure and organization of chromatin domains in eukaryotic chromosomes. DNA elements are necessary for the formation of chromatin domains and these elements also function to organize the domains within the three-dimensional context of the nucleus. Understanding long-range interactions between DNA elements and their role in the dynamics of chromatin domain is one focus of this proposal. The final goal is to understand how chromatin bound factors and DNA elements influence chromatin structure thereby affecting processes such as transcription. Adjacent chromatin domains in the nucleus often have antagonistic structural or functional properties and DNA insulator elements separate these domains. Promoters of specific genes often function as insulators in yeast and Drosophila and we have determined the broad outlines of the mechanisms by which these insulators function in yeast. In this grant we propose to investigate the changes that occur in chromatin domains as cells divide and to determine if similar mechanisms operate to separate chromatin domains in human cells. The research will involve use of molecular genetics and genomics, manipulation of chromatin using techniques like quantitative chromatin immunoprecipitations, and chromosome conformation capture and biochemical analysis. Understanding how the nucleus is organized should help us better manipulate processes for benefit in research. The results from this study will be useful in understanding the molecular basis of disease states that arise from mis-regulation of chromatin domains. The results will also enable researchers to develop new strategies for treating disease states by developing new molecular tools for gene therapy.