REGULATION OF RNA POLYMERASES DURING DEVELOPMENT

拟议研究的主要长期目标是确定 参与转录和RNA聚合酶调节的机制 在生长和发育的过渡期。 重点将是 放置在静止组织被诱导增殖的过渡区， 环境或荷尔蒙信号。 具体目标包括：（1） 确定核RNA聚合酶的亚基结构关系 在真核生物中（即，动物、植物和真菌）， 针对RNA聚合酶产生的亚单位特异性抗体作为探针。 RNA 聚合酶亚基蛋白印迹法将鉴定结构 这些酶在亚基水平的关系，并提供信息 通过进化来保护结构。 (2)以确定 RNA聚合酶亚基结构的改变是否在 在生长和发育过程中转录的调节。 这将 通过分析发育过程中的RNA聚合酶结构进行评价 使用亚基特异性抗体作为探针。 (3)使溶于 体外转录系统能够准确地从 来源，这将允许纯化的转录因子， 同质性 (4)利用现有的体外转录系统， 分析单个RNA聚合酶亚基的功能。 RNA聚合酶的亚基特异性抗体将促进这些 通过它们与RNA上的特定位点结合的能力进行研究 聚合酶，从而抑制转录中的特定过程。 (5)到 启动分离RNA聚合酶亚单位基因的实验， cDNA克隆到亚基mRNA。 RNA聚合酶亚基特异性 抗体将为cDNA的免疫学筛选提供探针 图书馆. 这些基因和cDNA克隆的分离将提供 用于研究RNA聚合酶合成调节的探针， 增长和发展转型。 RNA聚合酶的研究 结构、亚基功能和合成调节应提供 深入了解真核生物中的转录调控，特别是在某些情况下， 其中细胞被诱导到异常的增殖水平。

The major long range objective of the proposed research is to determine the mechanisms involved in the regulation of transcription and RNA polymerases during growth and developmental transitions. The major emphasis will be placed on transitions where quiescent tissues are induced to proliferate by environmental or hormonal signals. Specific objectives include: (1) To determine the subunit structural relationships of nuclear RNA polymerases among eukaryotic organisms (i.e., animal, plant, and fungal) by using subunit-specific antibodies raised against RNA polymerases as probes. RNA polymerase subunit protein blotting will identify the structural relationships of these enzymes at the subunit level and provide information on the conservation of the structures through evolution. (2) To determine whether the alteration in RNA polymerase subunit structures plays a role in the regulation of transcription during growth and development. This will be evaluated by analysis of RNA polymerase structures during development using subunit-specific antibodies as probes. (3) To develop soluble in vitro transcription systems that are capable of accurate transcription from sources that would allow the purification of transcription factors to homogeneity. (4) To use the available in vitro transcription systems for analyzing the function of individual RNA polymerase subunits. Subunit-specific antibodies to the RNA polymerases will facilitate these studies by their ability to bind to an block specific sites on the RNA polymerases, and thus inhibit a specific process in transcription. (5) To initiate experiments on the isolation of RNA polymerase subunit genes and cDNA clones to subunit mRNAs. The RNA polymerase subunit-specific antibodies will provide probes for immunological screening of cDNA libraries. The isolation of these genes and cDNA clones will provide probes for studying the regulation of RNA polymerase synthesis during growth and developmental transitions. These studies on RNA polymerase architecture, subunit function, and regulation of synthesis should provide insight into transcriptional regulation in eukaryotes especially in cases where cells are induced to abnormal levels of proliferation.