PAX GENE TRANSCRIPTION REGULATION IN EARLY EMBRYOGENESIS

含有配对盒(Pax)DNA结合域的蛋白质是关键 多种植物早期发育模式形成的调节因子 后生动物物种。这些基因的作用主要是从 功能突变的丧失，这已被证明是 人、小鼠、果蝇和果蝇的发育异常数量 秀丽隐杆线虫。尽管这些基因的基本作用是 似乎在后生动物的发育中，相对鲜为人知的是 它们转录调控的机制。这项工作 在这里提出的目的是为了理解转录调控 和开发海胆发育的两个方面 模型：(1)分离大量胚胎细胞核的能力 用于转录因子的分离，以及(2)检测的实用性 报告基因在大量发育中的胚胎中构建。这些 工具将被用来：(1)绘制海胆中的调控结合部位图 帕克斯同源基因。(2)确定这些位点在转录中的功能 控制，以及(3)识别和表征上游调控基因 在发育早期控制Pax的表达。的最终目标是 拟议的研究不仅定义了顺式约束的直接因素 以及允许受控表达Pax基因的机制。但 描述上游跨顺位监管网络的特点 最终达到它们的受控转录。鉴于明显的 已观察到的一些Pax函数的极端守恒性 从果蝇到人类的各种物种的基因。这些信息 从拟议的研究中获得的结果可能会有广泛的应用 了解脊椎动物中Pax表达的调控。

Proteins containing the Paired Box (Pax) DNA binding domain are key regulators of early developmental pattern formation in a wide variety of metazoan species. The role of these genes has chiefly been inferred from loss of function mutations, which have been shown to be the cause of a number of developmental anomalies in man, mouse, Drosophila and Caenorhabditis elegans. Despite the fundamental role that these genes appear to have in metazoan development, relatively little is yet known of the mechanisms by which their transcription is regulated. The work proposed here is directed at understanding the transcriptional regulation of Pax proteins and exploits two aspects of the sea urchin developmental model: (1) the ability to isolate immense quantities of embryonic nuclei for transcription factor isolation, and (2) the practicality of testing reporter gene constructs in large numbers of developing embryos. These tools will be used to: (1) map regulatory binding sites in a sea urchin Pax homolog. (2) determine the function of these sites in transcriptional control, and (3) identify and characterize upstream regulatory genes that control Pax expression during early development. The ultimate goal of the proposed research is to define not only the immediate cis-binding factors and mechanisms that allow for the controlled expression of Pax genes.but to characterize the upstream trans-cis regulatory networks that lead eventually to their controlled transcription. In view of the apparent extreme conservation of function that has been observed for a number of Pax genes in species ranging from Drosophila to humans. The information obtained from the proposed research is likely to have broad application towards understanding the control of Pax expression in vertebrates.