自吞噬（autophagy）是细胞内一种溶酶体依赖性的降解途径。自噬的细胞生物学过程可以概括为前自噬体的装配、自噬体膜的延伸、自噬体的形成、自噬体同溶酶体的融合、 自噬性溶酶体再生等5个步骤。 其中，自噬性溶酶体再生由申请人 发现并提出，在这一过程中，由自噬溶酶体膜形成的管状结构从 自噬溶酶体上伸出， 新溶酶体从这一管状结构上断裂形成。申请人通过大规模筛选工作鉴定了参与自噬性溶酶体再生的重要因子。在本项目中，我们基于前期研究工作，利用纯化的参与自噬性溶酶体再生的蛋白、小分子，人工膜或从细胞中纯化出的膜结构，在 体外重构系统中重构了自噬性溶酶体的出管过程，我们计划利用此系统，结合传统的细胞生物学手段， 深入研究自噬性溶酶体 的出管，货物选择，新溶酶体从自噬性溶酶体再生管上的断裂等过程的分子机制，体外重构在更可控的体系中研究生物学问题，能够更准确和深人和准确地研究自噬性溶酶体再生的分子机制 。

Autophagy is a lysosome-based degradation pathway. During autophagy, lysosomes fuse with autophagosomes to form autolysosomes. Following starvation-induced autophagy, nascent lysosomes are formed from autolysosomal membranes through an evolutionarily conserved cellular process, autophagic lysosome reformation (ALR), which is essential for maintaining lysosome homeostasis (Yu et al, Nature, 2010). During ALR, tubular structure (reformation tubules) extended from autolysosome, and nascent lysosomes are formed by fission of reformation tubules. Combining a screen of candidates identified through proteomic analysis of purified ALR tubules, and large-scale RNAi knockdown, we unveiled a tightly regulated molecular pathway that controls lysosome homeostasis (Rong et al, PNAS, 2011, Rong et al, Nature Cell Biology, 2012). In this project, We successfully set up an in vitro reconstitution system in which the tabulation of autolysosome is recapitulated. We will take advantage of this system to investigate the molecular mechanisms of ALR, including molecular mechanism governing autolysosome tabulation, cargo sorting and fission of nascent lysosome.