REGULATION OF RNA POLYMERASES DURING DEVELOPMENT

拟议研究的主要长期目标是确定 转录调控和RNA聚合酶的调控机制 在生长和发育过渡期间。主要重点将是 被放置在静止组织被诱导增殖的过渡上 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(即动物、植物和真菌)中使用 针对RNA聚合酶产生的亚基特异性抗体作为探针。核糖核酸 聚合酶亚单位蛋白印迹将鉴定结构 这些酶在亚单位水平上的关系，并提供信息 关于通过进化保护结构的问题。(二)确定 RNA聚合酶亚单位结构的改变是否在 转录调控生长发育过程中转录的调节这将是 通过分析发育过程中的RNA聚合酶结构进行评估 使用亚基特异性抗体作为探针。(3)发展成可溶于 体外转录系统能够准确转录来自 允许转录因子纯化的来源 同质性。(4)利用现有的体外转录系统进行 分析单个RNA聚合酶亚基的功能。 针对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)至 RNA聚合酶亚基基因分离和分离的初步实验 克隆到亚单位mRNAs的c DNA。RNA聚合酶亚基特异性 抗体将为免疫学筛选cDNAs提供探针 图书馆。这些基因和cdna克隆的分离将为 核糖核酸聚合酶合成调控研究的探讨 生长和发育的转变。关于RNA聚合酶的研究 体系结构、亚单位功能和合成规则应提供 对真核生物转录调控的洞察，特别是在案例中 细胞被诱导到不正常的增殖水平。

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.