FACTORS INVOLVED IN TRANSCRIPTION BY RNA POLYMERASE II

RNA 聚合酶 II 转录涉及的因素

• 批准号: 6986211
• 负责人: David H Price
• 金额: $ 39.99万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-01-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6986211
• 关键词: DNA directed RNA polymerase; Drosophilidae; Escherichia coli; RNA interference; cell cycle; confocal scanning microscopy; enzyme mechanism; immunofluorescence technique; methyltransferase; polyadenylate; posttranscriptional RNA processing; protein localization; protein purification; recombinant proteins; tissue /cell culture; transcription factor; transcription termination

DESCRIPTION (provided by applicant): The elongation phase of transcription by RNA polymerase II (Pol II) is highly regulated and intricately integrated into mRNA biosynthesis. In an effort to understand the factors involved and their mechanisms of action, studies are proposed that will help understand the role of P-TEFb and TTF2 in controlling transcription elongation and to what extent RNA processing is functionally coupled to transcription. I. P-TEFb plays a key role in regulating the critical checkpoint to productive elongation that occurs shortly after initiation. The kinase activity of P-TEFb is in turn uniquely regulated by reversible association with an RNP which contains 7SK RNA and HEXIM1 or HEXIM2. Parameters affecting the formation and dissociation of the P-TEFb/7SK/HEXIM complex will be examined using methods in vitro and in vivo. Details of the function of P-TEFb on specific genes and globally over all genes will be gathered using nuclear run-on reactions coupled to array methods. II. Transcription is shut off during mitosis and the RNA Pol I and Pol II termination activity of TTF2 is required for mitotic repression of transcription elongation. The role of TTF2 in mitotic transcription repression will be further studied and potential roles in transcription coupled repair, and interphase transcription will be examined. Also the potential role of TFIIF and other elongation factors in controlling TTF2 activity during transcription will be investigated. III. The processing reactions needed to generate functional mRNAs occur in part during elongation and are influenced by the transcription process. Newly developed assays will be used to elucidate the details of the function of the human capping enzyme and the cap methyl transferase in the context of transcription. Finally, using a newly developed in vitro system we will determine the extent to which polyadenylation is functionally coupled to transcription elongation and termination. Most of the studies described are aimed at understanding basic processes controlling gene expression, but P-TEFb plays a major role in HIV gene expression and may be a target for AIDS therapy. Also because a drug in clinical trials for cancer therapy, flavopiridol, targets P-TEFb, the proposed studies are important.

描述（由申请人提供）：RNA聚合酶II（POL II）的转录延伸阶段受到高度调节，并复杂地整合到mRNA生物合成中。为了了解所涉及的因素及其作用机制，提出了研究，该研究将有助于了解P-TEFB和TTF2在控制转录伸长率以及RNA处理在多大程度上与转录的多大程度的作用。 I. P-TEFB在调节启动后不久发生的生产伸长的关键检查点中起关键作用。 P-TEFB的激酶活性反过来又受到与含有7SK RNA和HEXIM1或HEXIM2的RNP的可逆关联的独特调节。影响P-TEFB/7SK/HEXIM复合物的形成和解离的参数将使用体外和体内方法检查。 P-TEFB对特定基因和全球所有基因的功能的详细信息将使用与阵列方法结合的核跑步反应来收集。 ii。在有丝分裂过程中关闭转录，RNA POL I和TTF2的POL II终止活性是有丝分裂抑制转录伸长所必需的。 TTF2在有丝分裂转录抑制中的作用将进一步研究，并在转录耦合修复中的潜在作用，并将检查相间转录。同样将研究TFIIF和其他伸长因子在转录过程中控制TTF2活性中的潜在作用。 iii。产生功能mRNA所需的处理反应部分发生在伸长率期间，并受转录过程的影响。新开发的测定法将用于阐明在转录背景下人类封盖酶和帽甲基转移酶功能的细节。最后，使用新开发的体外系统，我们将确定在功能上耦合转录伸长和终止的程度。所描述的大多数研究旨在理解控制基因表达的基本过程，但是P-TEFB在HIV基因表达中起主要作用，并且可能是AIDS治疗的靶标。同样，由于癌症治疗的临床试验中的药物，黄酮醇，靶向P-TEFB，因此拟议的研究很重要。