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-末端结构域（CTD）在协调RNA加工和其他共转录事件以实现适当基因表达中起关键作用。它由多个七肽重复序列（Y1 S2 P3 T4 S5 P6 S7）组成，这些重复序列从酵母到人类高度保守。CTD修饰的主要形式是七肽重复内的Ser 2和/或Ser 5的磷酸化。CTD磷酸化的水平和模式在转录周期期间由CTD激酶和磷酸酶的协同作用调节。大量的CTD磷酸化状态，也称为“CTD密码”，形成了将特定大分子复合物募集到转录聚合酶的基础。最近鉴定的新的磷酸化CTD相关蛋白已经将CTD的已知功能从介导RNA加工扩展到协调其他共转录事件，如染色质重塑。与CTD生物学的快速发展相比，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修饰酶和CTD介导的共转录复合物组装的结构和机制。在这个提议中，我们描述了结构，生化和功能的研究，以调查（1）的结构的SRI结构域的Set 2组蛋白甲基转移酶及其与磷酸CTD的相互作用;（2）之间的相互作用串联FF结构域的CA 150和磷酸CTD;（3）一个新的Ser 5特异性CTD磷酸酶，Ssu 72的结构和机制。公共卫生相关性（公共卫生相关性声明）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，已经开发出有效的手段来调节CTD磷酸化模式或募集CTD相关核因子，以增强病毒基因表达。我们对（磷酸化）CTD相关蛋白的结构理解的进展，它们与（磷酸化）CTD的相互作用，以及CTD修饰酶可能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。