Structural, Biochemical and Functional Studies of RNAPII CTD Interacting Proteins

RNAPII CTD 相互作用蛋白的结构、生化和功能研究

• 批准号: 7526527
• 负责人: Pei Zhou
• 金额: $ 31.2万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7526527
• 关键词: Affect; Binding; Biochemical; Biological Assay; C-terminal; Catalysis; Code; Complex; Consensus; Coupled; Development; Enzyme Kinetics; Enzymes; Eukaryota; Eukaryotic Cell; Event; Family; Gene Expression; Genetic Transcription; Goals; HIV-1; Health; Histone Deacetylase; Histone H3; Histones; Human; Human Biology; Individual; Knowledge; Literature; Macromolecular Complexes; Maps; Measurement; Mediating; Methylation; Modification; Molecular; Nuclear; Nuclear Protein; Nuclear Proteins; Numbers; Pattern; Peptides; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Play; Polymerase; Positioning Attribute; Process; Protein C; Protein Dephosphorylation; Protein Tyrosine Phosphatase; Proteins; Public Health; RNA Polymerase II; RNA Processing; RNA Splicing; Recruitment Activity; Relaxation; Role; Site-Directed Mutagenesis; Solutions; Staging; Structure; Testing; Transcription Elongation; Transcriptional Activation; Viral; Virus; Yeasts; base; chromatin remodeling; histone methyltransferase; human disease; improved; in vivo; insight; novel; structural biology; yeast genetics

DESCRIPTION (provided by applicant): The C-terminal domain (CTD) of RNA polymerase II (RNAPII) plays a pivotal role in orchestrating RNA processing and other co-transcriptional events to achieve proper gene expression. It consists of multiple heptad repeats (Y1S2P3T4S5P6S7) that are highly conserved from yeast to human. The predominant form of CTD modification is the phosphorylation of Ser2 and/or Ser5 within the heptad repeats. The level and pattern of CTD phosphorylation are regulated by the concerted action of CTD kinases and phosphatases during the transcription cycle. The vast number of CTD phosphorylation states, also known as the "CTD code," form the basis for recruitment of specific macromolecular complexes to the transcribing polymerase. Recent identification of novel phosphoCTD-associating proteins has expanded the known functions of the CTD from mediating RNA processing to coordinating other co-transcriptional events, such as chromatin remodeling. Compared to the rapid progress in CTD biology, the structural knowledge of CTD recognition by CTD- associating proteins and CTD-modifying enzymes is very limited. Our long term goal is to understand the structures and mechanisms of CTD-modifying enzymes and CTD-mediated assembly of co-transcriptional complexes. In this proposal, we describe structural, biochemical and functional studies to investigate (1) the structure of the SRI domain of the Set2 histone methyltransferase and its interaction with the phosphoCTD; (2) the interactions between tandem FF domains of CA150 and the phosphoCTD; (3) the structure and mechanism of a novel Ser5-specific CTD phosphatase, Ssu72. PUBLIC HEALTH RELEVANCE (Public Health Relevance Statement) Abnormal changes of CTD phosphorylation states and their associated processes have been connected to a variety of human diseases. A number of viruses, including HIV-1, have developed effective means to modulate the CTD phosphorylation pattern or recruit CTD-associating nuclear factors in order to enhance viral gene expression. Advances in our structural understanding of (phospho)CTD-associating proteins, their interactions with the (phospho)CTD, and CTD-modifying enzymes are likely to generate a significant impact on issues related to human disease and ultimately contribute to improving human health.

描述（由申请人提供）：RNA 聚合酶 II (RNAPII) 的 C 端结构域 (CTD) 在协调 RNA 加工和其他共转录事件以实现正确的基因表达方面发挥着关键作用。它由多个七肽重复序列 (Y1S2P3T4S5P6S7) 组成，这些重复序列从酵母到人类都高度保守。 CTD 修饰的主要形式是七肽重复序列中 Ser2 和/或 Ser5 的磷酸化。 CTD 磷酸化的水平和模式受到转录周期中 CTD 激酶和磷酸酶的协同作用的调节。大量的 CTD 磷酸化状态（也称为“CTD 代码”）构成了向转录聚合酶招募特定大分子复合物的基础。最近对新型磷酸CTD相关蛋白的鉴定将CTD的已知功能从介导RNA加工扩展到协调其他共转录事件，例如染色质重塑。与CTD生物学的快速进展相比，CTD相关蛋白和CTD修饰酶识别CTD的结构知识非常有限。我们的长期目标是了解 CTD 修饰酶的结构和机制以及 CTD 介导的共转录复合物组装。在本提案中，我们描述了结构、生化和功能研究，以研究 (1) Set2 组蛋白甲基转移酶的 SRI 结构域的结构及其与磷酸 CTD 的相互作用； (2) CA150串联FF结构域与磷酸CTD之间的相互作用； (3)新型Ser5特异性CTD磷酸酶Ssu72的结构和机制。公共健康相关性（公共健康相关性声明） CTD 磷酸化状态及其相关过程的异常变化与多种人类疾病有关。许多病毒，包括 HIV-1，已经开发出有效的方法来调节 CTD 磷酸化模式或招募 CTD 相关核因子，以增强病毒基因表达。我们对（磷酸）CTD 相关蛋白的结构理解、它们与（磷酸）CTD 的相互作用以及 CTD 修饰酶的进展可能会对与人类疾病相关的问题产生重大影响，并最终有助于改善人类健康。