PHOSPHORYLATION IN A DEVELOPMENTAL SIGNALING PATHWAY

发育信号通路中的磷酸化

• 批准号: 2690071
• 负责人: Steven Alexander Wasserman
• 金额: $ 24.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2690071
• 关键词: Drosophilidae; actin binding protein; biological signal transduction; cellular polarity; gene expression; gene interaction; gene mutation; genetic mapping; immunoprecipitation; in situ hybridization; laboratory rabbit; microinjections; phosphorylation; polymerase chain reaction; protein kinase; protein structure function; transcription factor; yeast two hybrid system

The long term goal of this research is to understand a signal transduction pathway that regulates programs of gene expression by controlling the subcellular localization of regulatory proteins. In the fruit fly Drosophila, intracellular transduction of a signal received at the surface of the embryo promotes nuclear translocation of the transcription factor Dorsal and thereby establishes the dorsoventral axis. In flies, plants, and mammals, homologous pathways direct nuclear import of related transcription factors as a critical step in immune or defensive responses. In addition to Dorsal, the Drosophila dorsoventral signaling pathway requires the transmembrane receptor Toll, the scaffolding protein Tube, the protein kinase Pelle, and the inhibitor Cactus. The fact that these five pathway components have been characterized at both the genetic and molecular level and can be assayed by embryo microinjection makes this system particularly amenable to experimental investigation. It is now possible, therefore, to address fundamental questions about the mechanism for signal transduction. The first two specific aims of this proposal will focus on the initiation of intracellular signaling. The potential function of the actin-binding protein Filamin as a link between Toll and Tube will be investigated in coimmunoprecipitation and genetic interaction studies. Additional experiments will address the mechanism and significance of Tube relocalization in response to Toll activation. The third and fourth aims will address how the signal is propagated. Autophosphorylation will be explored as a potential mechanism for Pelle regulation in vivo. Molecular and biochemical studies will also be carried out to determine which pathway proteins undergo Pelle-mediated phosphorylation in embryos. The fifth and final specific aim will focus on the spatial regulation of signaling within the syncytial environment of the blastoderm embryo. Given the conserved nature of the signaling pathway, the results of the proposed research should be of substantial interest with regard not only to pattern formation in the insect embryo, but also to signal transduction and immunity in a broad range of plants and animals.

这项研究的长期目标是了解信号 通过调节基因表达程序的转导途径 控制调节蛋白的亚细胞定位。 在 果蝇果蝇，接收的信号的细胞内转导 在胚胎的表面促进 转录因子背面，从而建立背腹 轴。 在苍蝇，植物和哺乳动物中，同源途径指导核 导入相关的转录因子作为免疫或 防御性反应。 除背侧，果蝇背腹信号传导途径 需要跨膜受体Toll，脚手架蛋白管， 蛋白激酶pelle和抑制剂仙人掌。这些事实 在遗传和 分子水平可以通过胚胎微注射来测定这一点 系统特别适合实验研究。 现在 因此，有可能解决有关的基本问题 信号转导机制。 该提案的前两个具体目标将重点放在 启动细胞内信号传导。 潜在功能 肌动蛋白结合蛋白丝作为收费和管之间的联系 在共免疫沉淀和遗传相互作用研究中进行了研究。 其他实验将解决 响应TOLL激活的管重新定位。 第三和 第四目标将解决信号传播的方式。 自磷酸化将作为Pelle的潜在机制探索 体内调节。 分子和生化研究也将是 进行以确定哪种途径蛋白经历了pelle介导的 胚胎中的磷酸化。 第五个也是最后的特定目标将集中 关于合成环境中信号传导的空间调节 胚胚胚。 鉴于信号通路的保守性质， 拟议的研究不仅应该具有很大的兴趣 在昆虫胚胎中形成图案，但也发出信号 在广泛的动植物中转导和免疫。