YEAST TRANSCRIPTIONAL ADAPTORS IN NUCLEOSOME ACETYLATION

这项拟议项目的总体目标是调查 真核转录调控与染色质的关系 结构，使用酿酒酵母作为这些研究的模式生物 学习。研究设计的重点是酵母纯化的多亚基 能够利用核小体作为底物的复合体 乙酰化。已提纯的两个配合物和部分 经鉴定含有转录适配子GCN5 (组蛋白乙酰转移酶)和ADA2。其中一个建筑群，佐贺， 还含有几种转录相关的SPT蛋白。对于 拟议的研究，佐贺复合体将在体外进行测试 与TATA结合蛋白(TBP)和激活剂的相互作用，两 激活转录中的关键成分。结构和 SAGA的接头和SPT亚基之间的功能关系 还将通过相互作用和乙酰化研究进行研究。在……里面 此外，佐贺成分在转录本身中的作用将是 通过制备接头和SPT突变体的复合体进行检查 将它们应用于体外激活的转录系统中 核小体DNA模板。这些研究旨在解决 真核基因表达调控的基本机制， 它们可能与复杂的人类疾病过程有关，例如 癌症。因为转录调控长期以来一直被认为是 在这些领域很重要，染色质被越来越多地发现 在其中扮演着重要的角色，拟议的项目可能会提出新的 通过提供对疾病更好的理解来治疗疾病的方法 调控途径。

The overall objective of this proposed project is to investigate the relationship between eukaryotic transcriptional regulation and chromatin structure, using Saccharomyces cerevisiae as a model organism for these studies. The research design focuses on yeast-purified multisubunit complexes that are capable of using nucleosomes as a substrate for acetylation. Two complexes that have been purified and partially characterized were shown to contain the transcriptional adaptors GCN5 (a histone acetyltransferase) and ADA2. One of these complexes, SAGA, also contains several transcriptionally relevant SPT proteins. For the proposed studies, the SAGA complex will be tested in vitro for interaction with the TATA-binding protein (TBP) and activators, two crucial components in activated transcription. The structural and functional relationship between the adaptor and SPT subunits of SAGA will also be investigated by interaction and acetylation studies. In addition, the roles of SAGA components in transcription itself will be examined by preparing complexes from adaptor and spt mutants and employing them in an in vitro activated transcription system with a nucleosomal DNA template. These studies are intended to address fundamental mechanisms of the regulation of eukaryotic gene expression, which likely have relevance to complex human disease processes such as cancer. Since transcriptional regulation has long been recognized as important in these areas, and chromatin is increasingly discovered to have a significant role therein, the proposed project may suggest novel approaches to disease treatment by providing a better understanding of regulatory pathways.