SOMA-TO-GERMLINE SIGNALING IN OOGENESIS IN DROSOPHILA

果蝇卵发生过程中的体细胞到种系信号传导

• 批准号: 6380133
• 负责人: STEPHEN M JACKSON
• 金额: $ 7.62万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6380133
• 关键词: Drosophilidae; biological signal transduction; cell type; cyclic AMP; cytoskeleton; developmental genetics; enzyme activity; oogenesis; phenotype; protein kinase A; protein kinase C; protein localization

The cut locus of D. melanogaster is an integral part of a communication pathway between two different cell types of the ovary. Loss of cut function in the somatic follicle cells interferes with these communications, disrupting the structural integrity of the germline-derived cells enclosed within the follicular epithelium We have shown by genetic analyses that mutations in cyclic-AMP dependent Protein kinase A (Pka-C1), agnostic, a gene that affects cAMP metabolism, and cappuccino and ovarian tumor, two genes that influence cytoskeletal function, interact genetically with cut mutations in this process. These results led us to hypothesize that cut activity in the follicle cells regulates a signaling pathway that impinges on the function of agnostic, cappuccino, and ovarian tumor in the germline cells. In this proposal, the function of these genes in this novel pathway is analyzed using a four-pronged approach. First, we will determine the mechanism responsible for the response of cappuccino and ovarian tumor to cut- and cAMP-mediated signaling by examining the expression, localization and phosphorylation state of these proteins when these pathways are disrupted. Second, we will determine the mechanism used by cut and agnostic in regulating cAMP levels and Pka-C activity in the germline by measuring cAMP levels, Pka-C catalytic activity, and agnostic expression levels when cut function is disrupted. Third, we will determine the importance of localized Protein kinase A activity in regulating the morphology of the germline cells by asking whether ectopic expression of PKA constructions or a dominant negative AKAP fragment can produce a multinucleate cell phenotype. Finally, we ask whether an A-kinase anchoring protein uncovered by a deficiency that interacts with cut mediates germ cell cytoskeletal integrity. Together, these experiments will determine how a communication pathway transmits a signal from one cell type through Protein kinase A to the cytoskeleton of another cell type.

黑腹果蝇的切割位点是卵巢两种不同细胞类型之间通讯途径的一个组成部分。体细胞滤泡细胞剪切功能的丧失会干扰这些通讯，破坏囊泡上皮内封闭的生殖系衍生细胞的结构完整性我们通过遗传分析表明，环磷酸腺苷依赖性蛋白激酶 A (Pka-C1) 发生突变，不可知论，一个影响cAMP代谢的基因，而卡布奇诺和卵巢肿瘤这两个影响细胞骨架功能的基因，在这个过程中与剪切突变发生遗传相互作用。这些结果使我们推测，卵泡细胞活性的降低调节了影响生殖细胞中不可知论、卡布奇诺和卵巢肿瘤功能的信号通路。在该提案中，使用四管齐下的方法分析了这些基因在这一新途径中的功能。首先，我们将通过检查当这些途径被破坏时这些蛋白质的表达、定位和磷酸化状态来确定卡布奇诺咖啡和卵巢肿瘤对 cut 和 cAMP 介导的信号传导的反应机制。其次，我们将通过测量 cut 功能被破坏时的 cAMP 水平、Pka-C 催化活性和不可知的表达水平来确定 cut 和不可知的调节种系中 cAMP 水平和 Pka-C 活性的机制。第三，我们将通过询问 PKA 结构的异位表达或显性失活 AKAP 片段是否可以产生多核细胞表型来确定局部蛋白激酶 A 活性在调节生殖细胞形态中的重要性。最后，我们询问由与切割相互作用的缺陷发现的 A-激酶锚定蛋白是否介导生殖细胞细胞骨架的完整性。这些实验将共同确定通讯途径如何通过蛋白激酶 A 将信号从一种细胞类型传递到另一种细胞类型的细胞骨架。