细胞自噬在生物的生长发育和疾病发生中具有重要作用，乙酰化修饰被报道参与调控细胞自噬的发生。申请人前期研究发现：家蚕自噬（ATG）蛋白发生去乙酰化修饰后出核，并诱导自噬；Rpd3为调控ATG蛋白出核的关键去乙酰酶。然而，酵母和哺乳动物中Rpd3同源蛋白相反地调控细胞自噬的发生，具体差异机制不详。申请人进一步检测发现，蜕皮激素（20E）信号诱导家蚕Rpd3的转录及其蛋白出核；Rpd3基因过表达诱导大部分Atg基因的表达和自噬发生。据此，本项目拟利用免疫沉淀结合蛋白质谱技术鉴定家蚕Rpd3存在的翻译后修饰方式和位点，解析该修饰方式和位点对Rpd3亚细胞定位和细胞自噬的影响；利用蛋白组学鉴定受Rpd3修饰、参与调控自噬发生的关键因子并进行功能验证，从而解析20E调控Rpd3参与调控细胞自噬的分子机制。本研究有利于阐释进化尺度中间物种的昆虫细胞自噬发生的分子机制，为高等动物的相关研究提供借鉴与参考。

Autophagy is essential for development and pivotal in disease occurrence in eukaryon. Acetylation on autophagy related (ATG) protein is reported to regulate autophagy occurrence. Previous work of the applicant showed that acetylation of ATG proteins facilitated their nuclear exportation and thus autophagy in Bombyx mori; moreover, deacetylase Rpd3 was revealed to be the key enzyme in regulating the translocation of ATG proteins. In addition, the molting hormone (20-hydroxyecdysone, 20E) signaling induced the transcription of Rpd3 and its nuclear exportation. However, the homologs of Rpd3 oppositely mediate autophagy in yeast and mammals, that the detailed different mechanism is unclear. The applicant further found that overexpression of Rpd3 induced most of all Atg genes expression and the occurrence of autophagy. Therefore, in this project, immunoprecipitation associated with mass-spectrum will be used to identify the post-translation modification types and sites of B. mori Rpd3, which would be verified for their roles in the subcellular localization of Rpd3 and autophagy. Protemics will be performed to identify the key Rpd3-deacetylated and interacted factors involved in the regulation of autophagy, for illustrating the mechanism underlying autophagy manipulated by 20E-regulated Rpd3. These researches will not only help to illustrate the molecular mechanism of autophagy during the species from intermediate evolutionary-scale such as insects, but also provide references for studies of autophagy in higher animals.