FUNCTIONAL SPECIFICITY OF THE DEFORMED PROTEIN

变形蛋白的功能特异性

基本信息

批准号：
2392450
负责人：
WILLIAM JAMES MCGINNIS
金额：
$ 20.04万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-04-01 至 1998-03-31
项目状态：
已结题

项目摘要

The goal of this research is to obtain a better mechanistic understanding of the factors underlying the functional specificity of Drosophila homeotic and mammalian Hox proteins, particularly those in the Deformed-like class. The core DNA sequence elements that are sufficient to confer Deformed specificity on a regulatory element in the context of an embryo are unknown. This proposal aims at the definition of such elements through mutagenesis studies on two Deformed autoregulatory enhancers (one specific for epidermal cells, and one for central nervous system cells). In addition, a human autoregulatory element that supplies maxillary segment-specific expression in Drosophila will be dissected to test whether it also contains Deformed response elements similar to those present in Drosophila enhancers. Using test of the function of Deformed/Ultrabithorax hybrid protein expression in Drosophila under heat shock promoter control, the targeting specificity of the Deformed and Ultrabithorax proteins will be analyzed. Affinity methods will be used to screen for proteins that interact with Deformed homeodomain proteins. As another approach to identify proteins that interact with, and contribute to the regulatory specificity of the Deformed protein, we will screen for mutations that enhance Deformed mutant phenotypes. Expression of some mouse and human Hox proteins in Drosophila causes gain-of-function mutations similar to the phenotypes caused by their Drosophila homologs. One simple hypothesis to explain this is that the mouse and human Hox proteins may conserve cross-regulatory relationships with extended members of the HOM/Hox system that may include the ems, Dll, otd, lab, and tsh genes. We will test this hypothesis using genetic epistasis tests and analysis of expression patterns in Hox protein expressing Drosophila embryos.

这项研究的目的是获得更好的机械理解果蝇功能特异性的因素同源性和哺乳动物HOX蛋白，尤其是那些类似变形的类。足够的核心DNA序列元素在监管元素上赋予变形的特异性胚胎是未知的。该建议旨在定义通过诱变研究对两个变形自动调节的元素增强剂（一种针对表皮细胞的特异性，一种用于中枢神经系统单元格）。此外，提供的人类自动调节元素提供果蝇中的上颌段特异性表达将剖析测试它是否还包含类似于该元件的变形响应元素存在于果蝇增强剂中。使用测试的功能在热量下，果蝇中的变形/超硫代术混合蛋白表达冲击启动子控制，变形的靶向特异性和将分析超稳定蛋白。将使用亲和力方法筛选与变形同源域蛋白相互作用的蛋白质。作为识别与与之相互作用的蛋白质的另一种方法有助于变形蛋白的调节特异性，我们将筛选突变增强变形突变表型的突变。表达果蝇原因中的一些小鼠和人Hox蛋白功能收益突变类似于由其作用的表型果蝇同源物。一个简单的假设可以解释这一点小鼠和人类HOX蛋白可以保留交叉调节关系 HOM/HOX系统的扩展成员可能包括EMS， DLL，OTD，LAB和TSH基因。我们将使用遗传来检验这一假设 HOX蛋白中表达模式的上毒测试和分析表达果蝇胚胎。