TRANSCRIPTION REGULATION BY ONCOGENES

哺乳动物细胞中RNA聚合酶II的转录调节将 在生物化学水平上进行研究。 分析这一问题的进展 基本的过程将被用来阐明分子的性质， 控制恶性生长。 通过序列激活转录- 特定的转录因子改变了基础复合物的结构 至少由IIA IID因素组成。 这件事的独特之处在于 将确定指定引发的增加速率的复合物。 转录因子与基础因子通过特异性 蛋白质接触 这些蛋白质信号的化学性质 将阐明和接触的基础因素的表面将是 鉴定 对酵母的分析结果表明，聚合酶II 与IIA IID复合物结合，作为含有其他基础酶的全酶， 因子和细胞成分。 类似全酶的证据 在哺乳动物系统中寻找。 Oct-1和Oct-2转录因子结合相同的八聚体序列 但在B细胞发育过程中具有不同的基本作用。 录病毒 载体将用于引入Oct-1和Oct-2的嵌合体，以确定 决定其在B细胞中独特作用的蛋白质信号。 一 序列特异性转录的合理设计方案 一种以显性方式调节特定内源性 基因被提出。 该方案依赖于共价连接的 序列特异性结构域以最大化亲和力和特异性。 的 技术将通过连接器的变化进行扩展和测试 连接序列特异性结构域的序列，结合的突变， 锌指结构域的特异性和其他DNA结合的融合 域. 这一方法将通过设计一个主导因素进行测试 其将刺激或抑制c-Myc和其它基因的转录。 许多癌基因和肿瘤抑制基因编码结合DNA的蛋白质 并调节转录。 体外和体内试验将用于 研究c-Myc、Rb-E2 F型复合物的转录活性， Wilms肿瘤因子类型因子。

Regulation of transcription by RNA polymerase II in mammalian cells will be studied at the biochemical level. Advances in the analysis of this fundamental process will be used to elucidate the molecular nature of control of malignant growth. Activation of transcription by sequence- specific transcription factors alters the structure of a basal complex composed of at least the IIA IID factors. The unique aspect of this complex which specifies an increase rate of initiation will be determined. Transcription factors interact with basal factors through specific protein-protein contacts. The chemical nature of these protein signals will be elucidated and the surfaces of the basal factors contacted will be identified. Results from the analysis of yeast suggest that polymerase II associates with the IIA IID complex as a holoenzyme containing other basal factors and cellular components. Evidence for a similar holoenzyme will be sought in mammalian systems. The Oct-1 and Oct-2 transcription factors bind the same octamer sequence but have different essential roles during B cell development. Retroviral vectors will be used to introduce chimeras of Oct-1 and Oct-2 to determine the protein signals which determine their unique roles in B cells. A protocol for the rational design of a sequence-specific transcription factor which will regulate in a dominant fashion a specific endogenous gene has been proposed. This protocol depends upon covalent linkage of sequence-specific domains to maximize affinity and specificity. The technology will be extended and tested by variation of the linker sequences joining the sequence-specific domains, mutation of the binding specificity of the zinc finger domains and fusion of other DNA binding domains. This methodology will be tested by designing a dominant factor which will stimulate or suppress transcription of c-Myc and other genes. Many oncogenes and tumor suppressor genes encode proteins which bind DNA and regulate transcription. In vitro and in vivo assays will be used to study the transcriptional activities of c-Myc, Rb-E2F type complexes and the Wilms' tumor factor-type factors.