Kinetic Mechanisms of Chromatin Remodeling and Transcription

项目摘要 真核基因表达的进展已经提供了一个全面的转录相关蛋白的列表， 它们的生物化学活性和结构与功能的关系，揭示了组蛋白的重要性 与序列特异性DNA结合协同作用的修饰和核小体重塑酶 转录因子，导致基因启动子处的转录稳定的染色质结构， 增强剂。然而，主要的挑战仍然是缺乏对时间尺度和动力学的了解， 表观遗传和转录蛋白作用于染色质底物。该提案将解决这些问题。 通过专注于染色质重塑和转录的定量动力学，使用最先进的 应用于活细胞和体外固定化染色质模板的单分子成像技术， 结合传统生物化学和酵母分子遗传学。预期的发现是 关键的反应中间体，事件的顺序，和限速步骤，将大大提高基本， 对天然染色质转录的机制理解，对真核生物的其他领域具有高度影响 DNA代谢

Project Summary Advances in eukaryotic gene expression have provided a comprehensive list of transcription-related proteins, their biochemical activities and structure-function relationships, and revealed the importance of histone modifications and nucleosome remodeling enzymes that cooperate with sequence-specific DNA binding transcription factors, resulting in a transcriptionally poised chromatin architecture at gene promoters and enhancers. However, major challenges remain in the lack of knowledge of the timescales and kinetics by which epigenetic and transcription proteins operate on chromatin substrates. This proposal will address these challenges by focusing on quantitative kinetics of chromatin remodeling and transcription, using state-of-the-art single-molecule imaging techniques applied to living cells and immobilized chromatin templates in vitro, combined with traditional biochemistry and yeast molecular genetics. Anticipated findings are the identification of key reaction intermediates, order of events, and rate-limiting steps that will dramatically advance basic, mechanistic understanding of transcription on native chromatin with high impact on other areas of eukaryotic DNA metabolism.