CONTROL OF LOCAL GROWTH BY EXPANDED GENE IN DROSOPHILA

通过扩展基因控制果蝇的局部生长

• 批准号: 2397401
• 负责人: PETER J BRYANT
• 金额: $ 12.69万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2397401
• 关键词: Drosophilidae; apical membrane; binding proteins; biological signal transduction; cell growth regulation; cell proliferation; developmental genetics; gene interaction; genetic regulation; imaginal disc; immunoprecipitation; membrane proteins; nucleic acid probes; protein structure function; tumor suppressor genes; western blottings; yeast two hybrid system

Thre overall goal of our laboratory is to elucidate the mechanisms controlling cell proliferation. This proposal concentrates on the Drosophila gene expanded (ex) which encodes a member of the protein 4.1 family of membrane-associated proteins. This gene is unusual in that its effect on cell proliferation is region-specific. Loss-of- function ex mutations cause an increase in the size of the wing, whereas general over-expression of the normal gene causes a decrease in size. Thus, appropriate levels of Ex expression are important for the local control of cell proliferation and final wing morphology. The predicted structure of Ex and the ex mutant phenotype suggest that Ex functions in a signaling pathway. This possibility is being tested by identifying genes and proteins that interact with Ex using both genetic and biochemical approaches. The yeast two-hybrid system has been used to screen Drosophila cDNA libraries for clones encoding Ex- binding proteins, and to test specific proteins that are already suspected to interact with Ex. Direct binding to Ex will be verified, and the genes encoding Ex-binding proteins will be identified and characterized by genetic and molecular methods. The domain in Ex responsible for its localization to the apical cell-membrane will be identified by expressing germline constructs encoding epitope-tagged domains of Ex, and determining their subcellular localization. The hypothesis that Ex acts as a linker molecule will be explored by examining the phenotypes induced by over-expression of these domains. The work will open up a new avenue of research into the mechanisms that control local differences in the rate and amount of cell proliferation in developing organs.

我们实验室的总体目标是阐明机制 控制细胞增殖。 该建议集中于 果蝇基因扩展（EX），该基因编码蛋白质的成员 4.1膜相关蛋白家族。 这个基因在 它对细胞增殖的影响是特异性的。 丧失 功能突变导致机翼的大小增加， 正常基因的一般过表达导致降低 大小。 因此，适当的EX表达水平对于 细胞增殖和最终机翼形态的局部控制。 EX和EX突变表型的预测结构表明 信号通路中的EX函数。 这种可能性正在测试 通过识别使用两者与EX相互作用的基因和蛋白质 遗传和生化方法。 酵母双杂交系统具有 用于筛选果蝇cDNA库的编码克隆 结合蛋白质，并测试已经已经存在的特定蛋白 怀疑与Ex互动。 直接绑定到EX将被验证， 并且将确定编码前结合蛋白的基因，并 以遗传和分子方法为特征。 ex中的域 负责其在顶端细胞膜上的本地化将是 通过表达编码表位标签的种系结构来识别 EX的域，并确定其亚细胞定位。 这 EX充当接头分子的假设将由 检查这些结构域过表达引起的表型。 这项工作将为机制开辟新的研究途径 控制局部细胞速率和量的局部差异 发展器官的扩散。