Dynamics of histone-DNA interaction

项目摘要 这项研究计划的长期目标是阐明组蛋白-DNA相互作用在基因中的作用 监管。核小体是真核生物基因组的基本包装单位，它设置了物理屏障。 获得DNA，并需要在各种核过程中至少部分地拆解，例如 转录延长和染色质重塑。这些过程的动力学路径和速率是 基因调控机制中的关键成分，与组蛋白-DNA的动力学密切相关 互动。阐明动态的组蛋白-DNA相互作用在基因调控中的意义是一个 是当前染色质研究的重要而活跃的课题。这个项目的目的是阐明如何自发 组蛋白-DNA相互作用的动力学有助于调节两个重要的核过程 沿DNA移位的大型酶复合体。目标过程是转录延伸，通过 RNA聚合酶II和INO80对细胞活力和健康至关重要的染色质重塑 扩散。该项目将揭示强大而动态的组蛋白-DNA相互作用是如何牵连到 基于三色单分子荧光共振能量转移调控这些过程 (SmFRET)和单分子亚毫秒运动分析。这些创新方法将使 了解允许这些酶进入并作用于核小体的分子过程 他们活动的最终产品达到了前所未有的深度。

Project Summary The long-term objective of this research program is to elucidate the roles of histone-DNA interactions in gene regulation. The nucleosome, the fundamental packing unit of the eukaryotic genome, imposes physical barriers to DNA access and needs to disassemble at least in part during various nuclear processes such as transcription elongation and chromatin remodeling. The kinetic pathways and rates of these processes are crucial components in gene regulation mechanisms and closely coupled to the dynamics of histone-DNA interactions. Elucidating the implications of dynamic histone-DNA interactions in gene regulation is an important and active subject of the current chromatin research. This project aims to elucidate how spontaneous dynamics of histone-DNA interactions contribute to regulating two important nuclear processes catalyzed by large enzyme complexes that translocate along DNA. The target processes are transcription elongation by RNA polymerase II and chromatin remodeling by INO80 both of which are essential for cell viability and healthy proliferation. The project will reveal how the strong yet dynamic histone-DNA interactions are implicated in regulating these processes based on three-color single-molecule fluorescence resonance energy transfer (smFRET) and single-molecule sub-ms motion analysis. These innovative approaches will enable an understanding of the molecular processes that allow these enzymes to access and act on nucleosomes and the end products of their activities at an unprecedented level of depth.