High-resolution structural and global signaling network identification of the NEPH-NEPHRIN protein complex (A03)

我们建立了一个多物种平台的互补转基因小鼠与floxed基因的Nphs 1，Neph 1，Neph 2和Neph 3，以及原鸡和果蝇模型系统，将结合高分辨率，大规模的系列三维EM重建和功能研究，以解开的结构和功能的NEPH-NEPHRIN复合物在狭缝隔膜（SD）。这将为以下工作提供基础：（1）关联SD的高分辨率三维结构。(2)分析所有NEPH-NEPHRIN家族成员在SD的发育、稳定性、动力学和维持中的确切作用;（3）通过在果蝇肾细胞中使用全基因组RNAi抑制剂筛选系统地解码NEPH-NEPHRIN复合物的全局信号网络。

We established a multispecies platform of complementary transgenic mice with floxed genes for Nphs1, Neph1, Neph2 and Neph3, as well as Gallus gallus and Drosophila model systems that will be combined with high-resolution, large-scale serial 3D EM reconstructions and functional studies to unravel the structure and function of the NEPH-NEPHRIN complex at the slit diaphragm (SD). This will provide the foundation to: (1) Correlate the high-resolution 3D structure of SDs. (2) Analyze the precise role of all NEPH-NEPHRIN family members for the development, stability, dynamics and maintenance of the SD, and (3) Systematically decode the global signaling network of the NEPH-NEPHRIN complex by using a genome-wide RNAi suppressor screen in Drosophila nephrocytes.