RNA POL II POLY-A SITE AND 3' TERMINATION

RNA POL II POLY-A 位点和 3 终止

• 批准号: 3300485
• 负责人: ERIK S FALCK-PEDERSEN
• 金额: $ 22.47万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-01-01 至 1994-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3300485
• 关键词: Adenoviridae; DNA directed RNA polymerase; RNA splicing; genetic mapping; genetic terminator element; hemoglobin F; messenger RNA; mutant; nucleic acid sequence; polyadenylate; protein signal sequence; site directed mutagenesis; transcription factor; transcription termination; virus genetics; virus infection mechanism

The ability to regulate the mRNA population is a key element of normal cell function, required for cell cycle regulation, cell differentiation and cell determination. Minor mRNA changes can have a major impact on cell function, possibly best represented by the effects of altered oncogene expression or hormone induction. Clearly, promoter activation effected by both initiation and repression functions is a dominant player in mRNA metabolism, but it is not the only level at which mRNA populations can be controlled. A growing number of transcription units are also under the influence of "postinitiation" control functions which include premature termination (c-myc, c-myb, Ad-mlp), alternative splicing and alternative polyadenylation (Ad-mlp, calcitonin/CGRP, muscle proteins) and transcription termination (Ig mu-delta, Ad-mlp. In addition to these nuclear events, in the cytoplasm cell specific control of mRNA 1/2 life is also becoming more and more apparent. The "postinitiation" regulation of a cells mRNA population is the general area of research my group is addressing. This proposal deals specifically with experiments designed to understand the mechanisms which operate to regulate two of these events, poly(A) choice in complex transcription units and termination of RNA polymerase II transcription units. One important aspect of postinitiation functions which is presently open to debate is whether the regulation of these events is mediated directly through the RNA polymerase II elongation complex or are these events (particularly splicing and polyadenylation) controlled at a point which is uncoupled from the transcription complex. This issue is also important to the control of transcription termination, since we have demonstrated the DNA sequence AATAAA (polyadenylation signal sequence) is a required cis element of 3' termination. This proposal is directly answering this synthesis (using reconstructed adenovirus vectors) and in vitro dissection of the biochemical process involved in controlling the 3' postinitiation events. Basic information which will be generated by these studies will include comparison of in vivo and in vitro poly (A) site utilization for several polyadenylation signal elements, relate the efficiency of poly (A) site utilization to transcription termination, identify the 3' consensus sequence required to inducer transcription complex displacement and finally, demonstrate how for the adenovirus major late transcription unit, the generation of varied preinitiation complexes at the mlp can influence elongation and processing events at the 3' end.

调节mRNA种群的能力是正常细胞的关键要素 细胞周期调节，细胞分化和细胞所需的功能 决心。 较小的mRNA变化可能会对细胞产生重大影响 功能，可能最能体现的癌基因的影响 表达或激素诱导。 显然，启动子激活由 启动和抑制功能都是mRNA中的主要参与者 代谢，但这不是mRNA种群的唯一水平 受控。 越来越多的转录单元在 “启动后”控制功能的影响，包括过早 终止（C-MYC，C-MYB，AD-MLP），替代剪接和替代方案 聚腺苷酸化（AD-MLP，降钙素/CGRP，肌肉蛋白）和 转录终止（Ig Mu-Delta，Ad-Mlp。 核事件，在mRNA 1/2寿命的细胞质细胞特异性控制中 也变得越来越明显。 对 细胞mRNA种群是我小组的研究的一般领域 寻址。 该建议专门针对旨在的实验 了解以调节其中两个事件的机制， poly（a）在复杂转录单元中的选择和RNA终止 聚合酶II转录单元。 启动后功能的一个重要方面，目前开放 辩论是这些事件的调节是否是直接介导的 通过RNA聚合酶II延伸复合物，还是这些事件 （尤其是剪接和聚腺苷酸化）在一个点控制的点 与转录复合物未偶联。 这个问题对于 转录终止的控制，因为我们已经证明了 DNA序列AATAAA（聚腺苷酸信号序列）是必需的顺式 3'终止的元素。 该提议直接回答这个问题 合成（使用重建的腺病毒载体）和体外解剖 控制3'引发后涉及的生化过程 事件。 这些研究将生成的基本信息将 包括比较体内和体外聚（a）位点利用率 几种聚腺苷酸信号元件，层次 站点利用转录终止，确定3'共识 诱导者转录复合物位移所需的顺序和 最后，演示如何用于腺病毒主要转录单元， MLP处的各种前启动复合物的产生会影响 3'结束的伸长和处理事件。