细胞自噬是真核细胞清除胞内冗余物质的基本途径，对维持包括人类在内的真核生物的健康具有不可替代的作用。降解囊泡"自噬体"的形成是这一通路的核心步骤，也是当前本领域研究的重点之一。我们在前期工作中成功改良了现有红色荧光蛋白，解决了RFP在本领域应用的缺陷。我们另外发现：自噬蛋白Atg1不仅在此前一般设想的自噬起始阶段发挥作用，它在自噬体形成后期另有不同的功能。在此基础上，我们一方面将系统的通过活细胞显微观测确定各主要Atg蛋白在PAS参与自噬体形成的时间区段；另一方面将以部分Atg蛋白从PAS解离的现象为切入点，对包括Atg1在内的多阶段作用/多功能的自噬蛋白的"结构-功能"基础进行探索。我们的研究将全面覆盖当前已知的主要Atg蛋白。通过综合利用活细胞观测、遗传、生化等手段，我们有望获得一个全新的Atg蛋白参与自噬体形成的时序框架，并对部分多功能Atg蛋白的作用机制提出独特的诠释。

Autophagy is a major pathway for the clearance of intracellular materials. It is indispensable for the health of eukaryotic organisms, including human beings. The formation of the sequestration vesicles, autophagosomes, is the central step in the pathway and a focus of current research. In our preliminary study, we successfully constructed a newly-designed red fluorescent protein (RFP) with much improved function, eliminating a major obstacle in the utilization of RFPs in our field. We further found that Atg1, which is generally considered to act early during autophagosome formation, plays a different role at a later stage. Based on these findings, we set out to establish the temporal order of the actions of major Atg proteins during autophagosome formation by systematic live cell imaging. Focusing on the dissociation of specific Atg proteins from the PAS, we will explore the structural basis for the multi-functionality phenomena among Atg proteins. Our analysis will cover all the major Atg proteins. By combining live cell imaging, genetic and biochemical approaches, we seek to build a brand new temporal framework for the roles of Atg proteins during autophagosome formation; our work will also offer a new perspective on the subject of "multi-role" Atg proteins.