Elucidation of Integrator's Function

描述（由申请人提供）： 项目摘要：RNA聚合酶II（RNAPII）的C末端结构域（CTD）是蛋白质编码基因转录调控和RNA加工的重要组成部分。大量的数据也表明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，它含有至少12个新的亚基，称为整合子，除了核心RNAPII亚基。两个整合子亚基显示出与切割和多聚腺苷酸化特异性因子（CPSF）复合物亚基的相似性。我们发现Integrator被募集到U1和U2 snRNA基因并介导其3 '端加工。整合子复合物在后生动物中是进化上保守的，并且直接与RNA聚合酶II大亚基的C末端结构域相互作用。 该提议集中于阐明整合子在控制转录和细胞增殖中的作用。这三个目标旨在提供完整的Integrator复合体物理和功能特性。Aim 1描述了将开始定义复合物中单个蛋白质功能的实验，并详细描述了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控中的作用扩展了这项工作。 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和生长控制的调节具有重要意义。核小RNA加工的异常调节可通过剪接体的失调导致信使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： 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和生长控制的调节具有重要意义。核小RNA加工的异常调节可通过剪接体的失调导致信使RNA加工缺陷，并可能导致细胞转化和肿瘤发生。

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.