CELLULAR REGULATION BY HOMOLOGOUS ONCOPROTEINS

本提案的总体目标是深入了解 两个密切相关的差异表达的机制 相关原癌基因产物cJun和Jun B。之一 肿瘤发生的工作模型是 显性作用与显性否定之间的表达 基因c-jun和jun B，导致a 可以提供恶性表型的一组基因。 到 为了实现这一建议的总体目标，我们的近期目标是 为了进一步了解细胞的激活， 介导jun B基因表达诱导的因子。 顺式元件和细胞因子的鉴定， 通过刺激介导jun B表达将有助于我们 了解这种蛋白质的差异行为，Jun B，从 2009年6月。 本提案的具体目标是：1.）到 在jun B 5 '调控区中定义顺式作用元件 参与TPA增强jun B基因的表达， 毛喉素2.）的情况。分离和表征反式作用因子 与jun B 5 '中定义的顺式作用元件结合， 监管区域。3.）第三章产生抗血清 由细菌表达的cDNA克隆产生的反式激活因子， 用它来阐明生化机制。4.）以确定 分子。事件导致激活新的 通过调节jun B基因诱导物鉴定的反式激活因子 表情 这些目标可以通过以下方式实现： 转染，CAT测定，足迹分析，凝胶阻滞， 蛋白质纯化，文库筛选，无细胞 转录-翻译I系统和免疫沉淀。 希望对jun B基因的深入研究 表达将导致更好地理解身份 Jun B在负性调节c-jun表达中的作用。 因此，实现这些目标应扩大我们的 同源癌蛋白对细胞调控的知识。

The overall aims of this proposal are to gain insight into the mechanisms involved in differential expression of two closely related proto-oncogene products, cJun and Jun B. One of the working models for oncogenesis is that of the imbalanced expression between the dominant acting and dominant negative genes, c-jun and jun B, leading to increased transcription of a set of genes which can provide the malignant phenotype. To achieve the overall aims of this proposal, our immediate goal is to further our understanding of the activation of cellular factor(s) which mediate the induction of jun B gene expression. The identification of cis-elements and cellular factors that mediate induction of jun B expression by stimuli will help us to understand the differential behavior of this protein, Jun B, from cJun. The specific objectives of this proposal are: 1.) To define cis-acting elements in the jun B 5'-regulatory region involved in enhanced expression of the jun B gene by TPA and forskolin. 2.) To isolate and characterize trans-acting factor(s) that bind to the defined cis-acting element in the jun B 5'- regulatory region. 3.) To generate antisera against transactivator produced by a bacterially expressed cDNA clone and use it to elucidate the biochemical mechanisms. 4.) To determine the molecular. events leading to the activation of a newly identified transactivator by inducers that regulate jun B gene expression. These objectives can be accomplished by transfection, CAT assays, footprint analysis, gel retardation, protein purification, library screening, cell-free transcription-translation i systems, and immunoprecipitation. It is expected that in-depth investigation of jun B gene expression will lead to a better understanding of the identity and function of Jun B in negatively regulating c-jun expression. Therefore accomplishment of these goals should extend our knowledge of cellular regulation by homologous oncoproteins.