COFACTORS AND FUNCTIONS OF HOMEODOMAIN REPRESSORS

在大多数其他人类中，基因转录的变化在癌症的进展中是重要的 疾病、衰老过程以及多细胞生物体的发育 各阶段。详细了解这些变化是如何调节的是两种诊断的基础 工具和干预策略。进一步的进步意味着新方法的前景，以及 提高现有方法的有效性。重要的是，许多问题仍然存在 涉及的基本流程。序列特异性DNA结合蛋白及其辅因子 Recruit to the DNA是转录的最重要的调控因子之一。其中最大的一家 蛋白质家族共享DNA结合的同源结构域基序。在果蝇中可用的工具使它 有可能在真实的活体环境中详细研究作用和相互作用的机制。这项建议 是研究这个大家庭中几个成员的行动机制，以解决以下基本问题 关于特定DNA靶点的识别是如何完成的，以及 对招募辅助因子来抑制转录的认识，对染色质变化的认识 调解调节，以及正在被调节的发展途径。使用组合 体内和体外方法，这些研究将提供更清楚的理解如何 蛋白质组合识别体内适当的靶点，以及由此产生的后果 对下游基因和途径调控的认识。该项目的具体目标是： 1)确定产生同源结构域蛋白的靶基因特异性的机制 积少成多。研究从DNA结合到抑制直接靶标的机制 吉恩·索普配对了。 2)分析含有锌指DNA结合的内相互作用蛋白的功能 结构域，并测试该伙伴是否有助于靶基因特异性或激活/抑制 作用于体内特定的靶点。 3)鉴定和分析含有同源结构域的抑制子的功能结合部位- 跳过了草率配对的靶基因。确定负责顺序操作的机制 发展过程中的压抑由先偶跳后积的草率组合而成。

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.