RNA POLYMERASE II AND THE TRANSCRIPTION COMPLEX

描述：(改编自申请者摘要)基因表达调控 在转录水平上是调节细胞的主要手段之一 生长和分化。异常的转录调控在一种 在肿瘤转化中的重要作用。尽管进行了大量的研究 关于转录调控机制的活动，研究人员是 才刚刚开始了解多种转录因子以及如何 它们激活和抑制特定的转录。目前很少有 了解RNA聚合酶II中亚基相互作用的细节 (RNAP II)本身，或这些亚单位如何与RNAP II相互作用并被 大量的转录因子。我们的目标是理解这个结构， 真核转录机制的功能和调节，与一个 强调RNAP II。Burgess博士将研究RNAP II亚单位组织 以及架构和定位与一般转录相互作用的位置 溶液中的因素和在最低限度上形成的预引发络合物中的因素 发起人制度。这些研究建立在RNA方面的经验和进展的基础上 聚合酶和转录因子的纯化，并在制备和 利用单抗对蛋白质进行定量和纯化并抑制特定功能 互动。新技术，最近在校长中开发 研究人员的实验室，将使他能够快速绘制交互领域 RNAP II亚基和因子。他最近开发了一种多功能系统，用于 在酵母中表达酵母RNAP II亚基，现在可以详细询问 关于RNAP II在体内组装和功能的问题。在……方面取得了很好的进展 他与罗杰·科恩伯格博士在酵母的三维晶体结构上的合作 RNAP II提供了一个强大的框架，在其上放置亚基和 因素交互作图结果。 伯吉斯博士建议继续他之前最有希望的工作 资助期，并与佩吉博士合作开展一个新项目 Farnham，谁分享了McArdle的地板，研究相互作用 人类转录因子E2F1的激活域(AD)及其其余部分 转录机制，并使用抗E2F1 AD的单链抗体， 干扰其在体外和体内的功能。这是一个长期目标 研究是利用特定聚合酶/转录的详细知识 干扰异常调控的因子相互作用设计试剂 对维持肿瘤状态至关重要的相互作用。

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.