COFACTORS AND FUNCTIONS OF HOMEODOMAIN REPRESSORS

基因转录的变化在癌症的进展中是重要的，在大多数其他人类中， 疾病、衰老过程以及多细胞生物的发育 阶段详细了解这些变化是如何调节的是诊断和治疗的基础。 工具和干预策略。进一步的进步有望产生新的方法， 提高现有方法的有效性。重要的是，许多问题仍然存在， 涉及的基本过程。序列特异性DNA结合蛋白及其辅助因子 DNA的补充是转录的最重要的调节器之一。其中最大的 蛋白质家族共享DNA结合同源结构域基序。果蝇中的工具使得 可以在真实的体内环境中详细研究作用和相互作用的机制。这项建议 是研究这个家庭的几个成员的行动机制，以解决基本问题， 仍然是关于如何识别特定的DNA靶位点是完成的，以及 识别辅因子的募集来抑制转录，识别染色质的变化， 介导调节，以及被调节的发育途径。使用组合 在体内和体外的方法，这些研究将提供一个更清晰的了解如何 蛋白质的组合在体内识别适当的靶位点， 识别下游基因和途径的调节。该项目的具体目标是： 1)确定同源结构域蛋白产生靶基因特异性的机制 雕刻。研究从DNA结合到抑制直接靶点的机制 吉恩·邋遢搭档。 2)分析含有锌指DNA结合的En相互作用蛋白的功能 结构域，并测试该配偶体是否有助于靶基因特异性或激活/抑制 在体内的特定靶位点上发挥作用。 3)为了鉴定和分析含有同源结构域的阻遏蛋白Even- 在靶基因中被遗漏了。确定负责顺序的机制 抑制在发展过程中的草率配对的第一次甚至跳过，然后Engrailed。

Changes in gene transcription are important in the progression of cancer, in most other human diseases, and in the aging process, as well as in the development of multicellular organisms at all stages. A detailed understanding of how such changes are regulated is the basis of both diagnostic tools and intervention strategies. Further advancement holds the promise of novel approaches, and of increased effectiveness of current approaches. Importantly, many questions remain about the fundamental processes involved. Sequence-specific DNA binding proteins and the cofactors that they recruit to the DNA are among the most important regulators of transcription. One of the largest such families of proteins share the DNA binding homeodomain motif. Tools available in Drosophila make it possible to study mechanisms of action and interaction in detail in a true in vivo context. This proposal is to study mechanisms of action of several members of this family, to address basic questions that remain about how recognition of specific DNA target sites is accomplished, and the consequences of that recognition for the recruitment of cofactors to repress transcription, for chromatin changes that mediate regulation, and for the developmental pathways that are being regulated. Using a combination of in vivo and in vitro approaches, these studies will provide a clearer understanding of how combinations of proteins recognize appropriate target sites in vivo, and the consequences of that recognition for the regulation of downstream genes and pathways. The Specific Aims of the project are: 1) To determine the mechanisms that generate target gene specificity for the homeodomain protein Engrailed. Investigate the mechanisms that lead from DNA binding to repression of the direct target gene sloppy paired. 2) To analyze the functions of an En-interacting protein that contains a zinc-finger DNA binding domain, and to test whether this partner contributes to target gene specificity, or to activation/repression function on specific target sites in vivo. 3) To identify and analyze functional binding sites for the homeodomain-containing represser Even- skipped in the target gene sloppy paired. Determine the mechanisms responsible for the sequential repression during development of sloppy paired by first Even-skipped and then Engrailed.