Mechanisms Regulating DNA Methylation Maintenance in Chromatin

项目摘要/摘要 DNA甲基化和组蛋白翻译后修饰(PTM)对染色质模板的影响 生物过程，而这些“表观遗传”标记在癌症中经常被异常调节。澄清： 因此，控制DNA甲基化和组蛋白PTM的机制对于改善我们的 理解表观遗传学在癌症中的作用。E3泛素连接酶uhrf1与 维持细胞DNA甲基化和解除uhrf1的调控与细胞增殖相关， 转移，以及对DNA损伤剂的超敏。这些最近uhrf1与 对DNA甲基化和癌症进展的调节表明这种蛋白可能是一种有利的治疗方法 目标。然而，uhrf1生物学的许多基本方面并不为人所知。这款K99/R00的首要目标是 因此，提出的建议是增进我们对uhrf1与染色质的相互作用的理解，以及 DNA甲基化遗传的调控。我的初步研究证实，甲基化的识别 Uhrf1串联Tudor结构域对组蛋白H3赖氨酸9的作用是其DNA甲基化维持所必需的 功能。这项提案中的研究将以这些发现为基础：1)定义多价染色质如何 参与驱动uhrf1的DNA甲基化维持功能，2)决定空间和 Uhrf1对DNA甲基化维持的时间贡献，3)定义uhrf1相互作用体，以及4) 开发化学探针作为研究uhrf1功能的工具。这项提议得到了强有力的指导。 和培训计划，为成功的独立研究生涯奠定坚实的基础 表观遗传程序的调控及其功能障碍如何导致癌症的发生和发展。

PROJECT SUMMARY/ABSTRACT DNA methylation and histone post-translational modifications (PTMs) elicit influence on chromatin-templated biological processes, and these `epigenetic' marks are often aberrantly regulated in cancers. Elucidation of mechanisms controlling DNA methylation and histone PTMs are therefore important to better our understanding of the role of epigenetics in cancer. The E3 ubiquitin ligase UHRF1 is genetically linked to the maintenance of cellular DNA methylation and deregulation of UHRF1 correlates with cell proliferation, metastasis, and hypersensitivity to DNA damaging agents. These recent connections of UHRF1 to the regulation of DNA methylation and cancer progression suggest this protein may be a favorable therapeutic target. Yet, many fundamental aspects of UHRF1 biology are not known. The overarching goal of this K99/R00 proposal is therefore to advance our understanding of the interaction of UHRF1 with chromatin and the regulation of DNA methylation inheritance. My preliminary studies established that recognition of methylated histone H3 at lysine 9 by the UHRF1 tandem Tudor domain is required for its DNA methylation maintenance function. Studies in this proposal will build upon these findings to: 1) define how multivalent chromatin engagement drives the DNA methylation maintenance function of UHRF1, 2) determine the spatial and temporal contribution of UHRF1 to DNA methylation maintenance, 3) define the UHRF1 interactome, and 4) develop chemical probes as tools to study UHRF1 function. This proposal is supported by a strong mentorship and training plan, building a solid foundation for a successful independent research career investigating the regulation of the epigenetic program and how its dysfunction leads to the initiation and progression of cancer.