RNA POLYMERASE II AND THE TRANSCRIPTION COMPLEX

描述：（改编自申请人的摘要）基因表达的调节 在转录水平上是调控细胞的主要手段之一， 生长和分化。异常的转录调控在 在肿瘤转化中起重要作用。尽管有大量的研究 活动的转录调控机制，研究人员是 我们才刚刚开始了解多种转录因子 它们激活和抑制特定的转录。目前， 已知RNA聚合酶II内亚基相互作用的细节 (RNAP II）自身或这些亚基如何与蛋白质相互作用并被蛋白质调节， 许多转录因子。我们的目标是了解结构， 功能和真核转录机制的调节， 重点是RNAP II。伯吉斯博士将研究RNAP II亚单位组织 并定位与一般转录相互作用的位点 因素都在溶液中，并在预引发复合物上形成的最小 启动子系统这些研究建立在RNA的经验和进展的基础上 聚合酶和转录因子纯化，以及在制备和 单克隆抗体用于定量和纯化蛋白质以及抑制特异性功能性抗体的用途 交互.新技术，最近开发的主要 研究人员的实验室，将使他能够迅速映射互动域上 RNAP II亚单位和因子。他最近开发了一种多功能系统， 在酵母中表达酵母RNAP II亚基，现在可以询问详细的 RNAP II在体内的组装和功能。在以下方面取得了很大进展： 他与Roger Kornberg博士合作研究酵母的三维晶体结构， RNAP II提供了一个强大的框架，在其上放置亚基， 因子交互作用映射结果。 伯吉斯博士建议继续他以前最有前途的工作 赠款期间，并与佩吉博士合作开展一个新的项目 法纳姆，谁分享麦卡德尔地板，研究的相互作用， 人转录因子E2 F 1的激活结构域（AD）与其余的 转录机制，并使用E2 F1 AD的单链抗体， 在体外和体内干扰其功能。长期目标是 研究是利用特定聚合酶/转录的详细知识 因子相互作用来设计干扰异常调节的代理人， 相互作用对维持肿瘤状态至关重要。

DESCRIPTION: (adapted from applicant's abstract) Regulation of gene expression at the level of transcription is one of the major means of regulating cell growth and differentiation. Abnormal transcriptional regulation plays an important role in neoplastic transformation. Despite a great amount of research activity on the mechanisms of transcriptional regulation, the investigator is only beginning to get a picture of the multiple transcription factors and how they activate and repress specific transcription. Very little is presently known about the details of the subunit interactions within RNA polymerase II (RNAP II) itself or how these subunits interact with and are modulated by the numerous transcription factors. The goal is to understand the structure, function, and regulation of the eukaryotic transcription machinery, with an emphasis on RNAP II. Dr. Burgess will study the RNAP II subunit organization and architecture and locate sites of interaction with general transcription factors both in solution and in pre-initiation complexes formed on a minimal promoter system. These studies build on experience and progress in RNA polymerase and transcription factor purification, and in the preparation and use of MAbs to quantify and purify proteins and to inhibit specific functional interactions. New techniques, recently developed in the principal investigator's laboratory, will allow him to rapidly map interaction domains on RNAP II subunits and factors. He has recently developed a versatile system for expressing yeast RNAP II subunits in yeast, that now allows to ask detailed questions about RNAP II assembly and function in vivo. Excellent progress in his collaboration with Dr. Roger Kornberg on the 3-D crystal structure of yeast RNAP II provides with a strong framework upon which to place the subunit and factor interaction mapping results. Dr. Burgess proposes to continue the most promising work from his previous grant period and to undertake a new project in collaboration with Dr. Peggy Farnham, who shares the floor of McArdle, to study the interaction of the activation domain (AD) of hun1an transcription factor E2F 1 with the rest of the transcription machinery and, using single chain antibodies to E2Fl AD, to interfere with its function in vitro and in vivo. A long-term goal of this research is to use the detailed knowledge of specific polymerase/transcription factor interactions to design agents that interfere with abnormal regulatory interactions crucial to maintaining the neoplastic state.