STRUCTURAL CHARACTERIZATION OF THE TAF1-TAF7 TFIID SUBCOMPLEX

TAF1-TAF7 TFIID 子复合物的结构表征

• 批准号: 8361716
• 负责人: JOHN D BAXTER
• 金额: $ 3.84万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361716
• 关键词: Biochemical; C-terminal; Catalytic Domain; Code; Complex; Escherichia coli; Eukaryota; Family member; Funding; Gene Expression Regulation; Genes; Grant; Histones; National Center for Research Resources; Nucleic Acid Regulatory Sequences; Phosphotransferases; Play; Principal Investigator; Process; Proteins; RNA Polymerase II; Research; Research Infrastructure; Resources; Role; Sequence Homology; Site; Source; Structure; TAF1 gene; TAF7 gene; Transcription Factor TFIID; Transferase; United States National Institutes of Health; cost; human BDP1 protein; human TAF1 protein; human TAF5 protein; interest; promoter; structural biology

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The transcription factor TFIID, composed of the TATA box binding protein (TBP) and 14 TBP associated factors (TAFs), plays a key role in regulation of gene expression by RNA Polymerase II. TFIID plays a critical role as a core promoter recognition factor that begins the process of assembling the transcriptional apparatus at the start site of protein coding genes in eukaryotes. A variety of biochemical studies of TFIID complexes have suggested that the largest subunit of the complex, TAF1, possesses histone acetyl-transferase and kinase activities that may play critical roles in the regulation of gene expression from TFIID dependent promoters. While these activities have been postulated, TAF1 domains possess no sequence homology with any known kinase or acetyl-transferase family members. Although some TAF1 domains and functions have been characterized, the largest C-terminal region of the TAF1 molecule identified as the likely enzymatic domain remains structurally unknown. Consequently we are interested in obtaining structural information for the TAF1 subunit of TFIID. We have previously identified a stable subcomplex of TFIID that consists of the putative catalytic region of the TAF1 subunit and the regulatory region of TAF7 and have determined conditions for the expression and purification of this complex from E. coli. We hope to determine the structure of the TAF1-TAF7 subcomplex of TFIID crystallographically and believe this will provide the first structural glimpse into the organization and assembly of the preinitiation complex.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 转录因子 TFIID 由 TATA 盒结合蛋白 (TBP) 和 14 个 TBP 相关因子 (TAF) 组成，在 RNA 聚合酶 II 基因表达调节中发挥关键作用。 TFIID 作为核心启动子识别因子发挥着关键作用，它启动了真核生物中蛋白质编码基因起始位点的转录装置组装过程。 TFIID 复合物的各种生化研究表明，该复合物最大的亚基 TAF1 具有组蛋白乙酰转移酶和激酶活性，可能在调节 TFIID 依赖性启动子的基因表达中发挥关键作用。虽然这些活性已被假定，但 TAF1 结构域与任何已知的激酶或乙酰转移酶家族成员不具有序列同源性。尽管一些 TAF1 结构域和功能已被表征，但被确定为可能的酶结构域的 TAF1 分子的最大 C 末端区域在结构上仍然未知。因此，我们有兴趣获得 TFIID 的 TAF1 亚基的结构信息。我们之前已经鉴定了 TFIID 的稳定亚复合体，它由 TAF1 亚基的假定催化区和 TAF7 的调节区组成，并确定了从大肠杆菌中表达和纯化该复合体的条件。我们希望从晶体学上确定 TFIID 的 TAF1-TAF7 子复合物的结构，并相信这将为预引发复合物的组织和组装提供第一个结构概览。