MOLECULAR ANALYSIS OF TRANSCRIPTIONAL REPRESSION

这个项目的长期目标是确定分子的特征 真核转录抑制因子调控基因的机制 表情。转录调控是关键方面的基础 许多生物过程，包括发育、免疫反应、 病理状态，如癌症和动态平衡的生理学 和适应。而真核生物中的转录激活是 开始从机械论的角度理解，消极的监管 人们对抑制物基因的了解较少。 这项研究将使用互补的生化和遗传方法来 确定果蝇Knirps阻遏蛋白的作用机制 调节转录。果蝇就是这样一个很好的模型系统 研究是因为广泛的遗传学，先进的工具 可用于产生转基因动物，以及已建立的系统 用于体外转录。克尼普斯抑制者，是 核受体超家族转录因子，发挥重要作用 在早期胚胎发育中起重要作用。初步工作表明， Knirps可以在短距离内抑制增强剂和基础启动子。 这一活动是一个重要的抑制因子家族的特征。 果蝇，很可能代表了其他动物的一种常见的压抑模式 真核生物。 我们将确定Knirps蛋白在体内对抑制至关重要的区域 使用两种新的果蝇胚胎分析方法。这些遗传信息将会 补体的体外研究，包括体外转录分析， 它将提供对Knirps活动的机械性见解，并指导 结合研究，将确定可能与之相互作用的蛋白质 克尼普斯调停镇压。我们还将确定Knirps-Interactive 利用酵母双杂交技术筛选蛋白质，并评价候选靶点 利用遗传和生化信息开发的蛋白质 其他化验。 很明显，果蝇和果蝇体内的基本回路调节基因 脊椎动物是高度保守的。对压抑的分析对于 了解从发育到癌症的各种生物过程。 例如，像视网膜母细胞瘤这样的转录抑制因子， 肾母细胞瘤和MDM2蛋白是人类肿瘤发生的重要因素。 该项目将采用多学科方法来提供 关于这种重要基因形式的详细机制信息 监管。

The long-term objective of this project is to characterize the molecular mechanisms by which eukaryotic transcriptional repressors control gene expression. Transcriptional regulation underlies crucial aspects of many biological processes, including development, immune responses, pathological states such as cancer, and the physiology of homeostasis and adaptation. While transcriptional activation in eukaryotes is beginning to be understood in mechanistic terms, negative regulation of genes by repressors is less well understood. This study will use complementary biochemical and genetic approaches to determine the mechanism by which the Drosophila knirps repressor protein regulates transcription. Drosophila is a good model system for such studies because of the extensive genetics, the sophisticated tools available for generating transgenic animals, and the established systems for in vitro transcription. The knirps repressor, a member of the nuclear receptor superfamily of transcription factors, plays an important role in early embryogenesis. Initial work indicates that knirps can repress enhancers and basal promoters from a short range. This activity is characteristic of an important family of repressors in Drosophila, and likely represents a common mode of repression in other eukaryotes. We will define regions of knirps protein vital for repression in vivo using two novel Drosophila embryo assays. This genetic information will complement in vitro studies, including in vitro transcriptional assays, which will provide mechanistic insights into knirps activity, and direct binding studies, which will identify proteins which may interact with knirps to mediate repression. We will also identify knirps-interacting proteins using a yeast two-hybrid screen, and evaluate candidate target proteins using genetic and biochemical information developed from the other assays. It is clear that the basic circuitry regulating genes in Drosophila and vertebrates is highly conserved. Analysis of repression is crucial for understanding biological processes ranging from development to cancer. For instance, transcriptional repressors such as the retinoblastoma, Wilm's tumor, and MDM2 proteins are important factors in human cancers. This project will employ a multidisciplinary approach to provide detailed mechanistic information about this important form of gene regulation.