Elucidation of Integrator's Function

积分器功能说明

• 批准号: 8308483
• 负责人: RAMIN SHIEKHATTAR
• 金额: $ 38.65万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8308483
• 关键词: Biochemical; Boxing; C-terminal; Cell Cycle Regulation; Cell Line; Cell Proliferation; Cis-Acting Sequence; Cleavage And Polyadenylation Specificity Factor; Code; Complex; Cytoplasm; DDX26 gene; Data; Functional RNA; Gene Expression Regulation; Genes; Genetic Transcription; Growth; In Vitro; Individual; Lead; Malignant Epithelial Cell; Mediating; Messenger RNA; Molecular; Names; Non-Small-Cell Lung Carcinoma; Nuclear; Nucleotides; Pattern; Phosphorylation; Play; Process; Prostate carcinoma; Proteins; RNA 3' End Processing; RNA Polymerase II; RNA Precursors; RNA Processing; RNA polymerase II largest subunit; Recombinants; Recruitment Activity; Regulation; Role; Seminal; Small Nuclear RNA; Spliceosomes; Testing; Transcript; Transcription Process; Transcriptional Regulation; Tumor Suppressor Genes; Tumor Suppressor Proteins; U1 small nuclear RNA; U2 small nuclear RNA; Work; design; in vivo; insight; mRNA Precursor; metaplastic cell transformation; novel; polypeptide; promoter; protein complex; public health relevance; reconstitution; research study; tumorigenesis

DESCRIPTION (provided by applicant): Project Summary: The C-terminal domain (CTD) of RNA polymerase II (RNAPII) is an essential component of transcriptional regulation and RNA processing of protein coding genes. A large body of data also implicates the CTD in the transcription and processing of RNAPII-mediated small nuclear RNA (snRNA) genes. However, the identity of the complex(es) that associate with the CTD and mediate the processing of snRNA genes have remained elusive. We describe an RNA polymerase II complex that contains at least twelve novel subunits, termed the Integrator, in addition to core RNAPII subunits. Two of the Integrator subunits display similarities to the subunits of the cleavage and polyadenylation specificity factor (CPSF) complex. We show that Integrator is recruited to the U1 and U2 snRNA genes and mediates their 3'-end processing. The Integrator complex is evolutionarily conserved in metazoans and directly interacts with the C- terminal domain of the RNA polymerase II large subunit. This proposal is focused at elucidating the role of the Integrator, in control of transcription and cellular proliferation. The three Aims are designed to provide a thorough physical and functional characterization of the Integrator complex. Aim1 describes experiments that will begin to define the function of the individual proteins in the complex and details the functional reconstitution of Integrator. Aim2 describes a detailed functional analysis of Integrator in transcriptional regulation of snRNA genes and extend the work to analysis of other non-coding RNA and RNAPII-mediated genes. Aim3 extends the work by analyzing the role of the complex and its individual subunits in oncogenesis and cell cycle regulation. Elucidation of the mechanism through which small nuclear RNAs are transcribed and processed to mature RNAs is of seminal importance in understanding regulation of gene expression and growth control. Aberrant regulation of small nuclear RNA processing could result in defective messenger RNA processing through deregulation of splicesome and may lead to cellular transformation and oncogenesis. PUBLIC HEALTH RELEVANCE: Public Health Relevance Elucidation of the mechanism through which small nuclear RNAs are transcribed and processed to mature RNAs is of seminal importance in understanding regulation of gene expression and growth control. Aberrant regulation of small nuclear RNA processing could result in defective messenger RNA processing through deregulation of splicesome and may lead to cellular transformation and oncogenesis.

描述（由申请人提供）： 项目摘要：RNA 聚合酶 II (RNAPII) 的 C 端结构域 (CTD) 是蛋白质编码基因的转录调控和 RNA 加工的重要组成部分。大量数据还表明 CTD 参与 RNAPII 介导的小核 RNA (snRNA) 基因的转录和加工。然而，与 CTD 相关并介导 snRNA 基因加工的复合物的身份仍然难以捉摸。我们描述了一种 RNA 聚合酶 II 复合物，除了核心 RNAPII 亚基外，它还包含至少 12 个新亚基，称为整合器。 Integrator 亚基中的两个与裂解和聚腺苷酸化特异性因子 (CPSF) 复合物的亚基相似。我们表明 Integrator 被招募到 U1 和 U2 snRNA 基因并介导它们的 3' 端加工。 Integrator 复合体在后生动物中在进化上是保守的，并且直接与 RNA 聚合酶 II 大亚基的 C 端结构域相互作用。 该提案的重点是阐明整合器在控制转录和细胞增殖中的作用。这三个目标旨在提供积分器综合体的彻底的物理和功能特征。 Aim1 描述了将开始定义复合物中各个蛋白质功能的实验，并详细介绍了 Integrator 的功能重建。 Aim2 描述了 Integrator 在 snRNA 基因转录调控中的详细功能分析，并将工作扩展到其他非编码 RNA 和 RNAPII 介导的基因的分析。 Aim3 通过分析复合物及其单个亚基在肿瘤发生和细胞周期调节中的作用来扩展工作。 阐明小核 RNA 转录和加工成成熟 RNA 的机制对于理解基因表达调控和生长控制具有重要意义。小核 RNA 加工的异常调节可能会通过剪接体的失调而导致信使 RNA 加工缺陷，并可能导致细胞转化和肿瘤发生。 公共卫生相关性： 公共卫生相关性 阐明小核 RNA 转录和加工成成熟 RNA 的机制对于理解基因表达调控和生长控制具有重要意义。小核 RNA 加工的异常调节可能会通过剪接体的失调而导致信使 RNA 加工缺陷，并可能导致细胞转化和肿瘤发生。