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）在编排RNA处理和其他共转录事件方面起着关键作用，以实现适当的基因表达。它由多个重复序列（Y1S2P3T4S5P6S7）组成，它们从酵母到人都高度保守。 CTD修饰的主要形式是七体重复序列中Ser2和/或Ser5的磷酸化。 CTD磷酸化的水平和模式受转录周期中CTD激酶和磷酸酶的一致作用调节。大量的CTD磷酸化状态，也称为“ CTD代码”，构成了将特定的大分子复合物募集到转录聚合酶的基础。新型磷酸化促进蛋白的最新鉴定已将CTD的已知功能从中介RNA加工到协调其他共转录事件，例如染色质重塑。与CTD生物学的快速进步相比，CTD-关联蛋白质和CTD改良酶对CTD识别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和共转录复合物的CTD介导的组装。在该建议中，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化的相互作用； （2）CA150的串联FF结构域与磷酸化的串联FF域之间的相互作用； （3）新型Ser5特异性CTD磷酸酶SSU72的结构和机制。公共卫生相关性（公共卫生相关声明）CTD磷酸化状态及其相关过程的异常变化已与各种人类疾病有关。包括HIV-1在内的许多病毒已经开发出有效的手段来调节CTD磷酸化模式或募集CTD缔解核因子，以增强病毒基因的表达。我们对（磷）CTD缔解蛋白的结构理解，其与（磷）CTD的相互作用以及CTD修饰酶的相互作用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。