RNA POL II POLY-A SITE AND 3' TERMINATION

调节mRNA群的能力是正常细胞的一个关键因素 细胞周期调节、细胞分化和细胞分化所必需的功能 决心。微小的信使核糖核酸变化可能对细胞产生重大影响 功能，可能最好的代表是癌基因改变的影响 表达或激素诱导。显然，启动子的激活是由 启动和抑制功能都是信使核糖核酸的主要参与者。 新陈代谢，但这不是唯一的水平，信使核糖核酸种群可以 控制住了。越来越多的转录单位也在 包括早产在内的“启动后”控制功能的影响 末端(c-myc、c-myb、Ad-MLP)、选择性剪接和选择性 多聚腺苷(Ad-MLP、降钙素/降钙素基因相关肽、肌肉蛋白)和 转录终止(Ig Mu-Delta，Ad-MLP.除了这些之外， 核事件中，在细胞质细胞中特异性控制的mRNA 1/2寿命是 也变得越来越明显。“启动后”的规则 细胞信使核糖核酸种群是我的团队研究的总体领域 地址。这项提案专门涉及旨在 了解调节其中两个事件的机制， 复杂转录单位中的Poly(A)选择与RNA的终止 聚合酶II转录单位。 印心后功能的一个重要方面目前是开放的 争论的焦点是对这些事件的监管是否直接调解 通过RNA聚合酶II伸长复合体还是这些事件 (尤其是剪接和聚腺苷酸化)控制在一个点上 与转录复合体解偶联。这个问题对 转录终止的控制，因为我们已经演示了 DNA序列AATAAA(聚腺苷酸化信号序列)是必需的顺式序列 3‘端接的元素。这项提议直接回答了这一点 合成(使用重组腺病毒载体)和体外解剖 参与控制3‘后启动的生化过程 事件。这些研究将产生的基本信息将 包括体内和体外Poly(A)位点利用的比较 几个多聚腺苷化信号元件，关系到聚(A)的效率 站点利用到转录终止，确定3‘共识 诱导转录复合体置换所需的序列和 最后，演示对腺病毒主要的晚期转录单位， 在MLP处各种预引发复合体的产生可能会影响 3‘端的伸长和加工事件。

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.