WASH蛋白作为WASP家族新发现的成员，其功能与调控机制的研究成为该领域的重要科学问题。合作者课题组发现WASH蛋白调控细胞自噬的发生，并初步分离和鉴定了一个新的WASH蛋白复合体，证实了这个复合体参与自噬调控。该延续合作课题将获取均一的生理状态的WASH蛋白复合体，探明这个复合体对自噬发生的调控作用。测定WASH复合体中亚复合物的晶体结构，分析WASH蛋白复合体的结构特征，探明WASH蛋白复合体的调控自噬的组装机制。获得高质量的生理性的WASH蛋白复合体样本，利用冷冻电镜技术对WASH复合体进行三维结构重构，测定生理性WASH复合体的三维结构，结合亚复合物的晶体结构信息，以期揭示WASH复合体调控自噬的功能与结构关系。并将对WASH蛋白复合体的结构信息进行功能验证，进一步核实WASH蛋白复合体发挥调控自噬作用的分子机制。该研究将加深对自噬发生机制的认识，提高我们在该领域的研究水平。

WASH is a new member of the WASP family. Understanding the function and regulation of WASH and its protein complexes has become an issue of great importance in the field. The collaborator’s laboratory at the Institute of Biophysics, CAS, discovered recently that WASH plays a role in autophagy initiation, and produced preliminary evidence for a new 600-kD WASH protein complex that is involved in the regulation of autophagy. The proposed project will focus on obtaining homogeneous samples of this WASH complex and characterizingin detail itsregulatory role in autophagy. We will also determine the crystal structures of the individual proteins and their sub-complexes of the WASH complex, analyze the structural features of the complex, and define the assembly mechanism of this autophagy machinery. In addition, we will utilize cryo-EM to reconstruct the 3-D structure of the entire WASH protein complex, combine the EM envelope with the crystal structure information, and carry out structure-based mutagenesis and functional studies to gain a deeper understandingof the structure and functionof the WASH complex. Overall, this project will greatly improve our knowledge on autophagy regulation,elevate our research and increase our international competitiveness in this area.