利用蛋白质组学方法研究N末端乙酰转移酶在线虫发育繁殖中的作用

Protein N-terminal acetylation is among the most common modifications. Recently it was discovered that N-terminal acetylation plays important roles in many biological processes， e.g., ubiquitin/proteasome pathway, apoptosis, protein interaction and subcellular localization and so on. Today, it is well recognized as a multifunctional regulator, and its function varies for different proteins in different circumstances. A systematic proteome scale study will help discover its new biological functions. Recently we developed a highly efficient proteomics approach for N-terminal acetylation study, which is composed of the purification using CNBr activated sepharose 4B and the identification using high mass accuracy mass spectrometer（Zhang, 2009; 2010; 2011; 2013）. Furthermore, we observed the severe developmental delay and birth reduction in C. elegans when the expression of NATs is inhibited using RNAi technique. The results suggest that N-terminal acetylation plays an important role during C. elegans development and reproduction. Taking advantage of the approach we developed, we plan to carry out a systematic study of the functions of N-terminal acetyltransferases (NATs) in the development and reproduction using C. elegans. We are going to systematically identify the large-scale N-terminal acetylome, elucidate the precise substrates for different NATs, screen out the key N-terminal acetylation sites relating to development and reproduction, and thus discover its regulation mechanism in development and reproduction. By achieving these purposes, we could extend our knowledge on protein N-terminal acetylation and may provide a new approach to study development and reproduction.

蛋白质N末端乙酰化是一种广泛存在的修饰方式，其相关的生物学功能研究最近才有所进展，主要参与泛素化降解和调节细胞凋亡等生物学过程，然而其更深入的生物学功能还有待进一步研究。通过系统的蛋白质组学研究将有助于发现其更多重要的生物学功能并揭示其作用的分子机制。本课题组在最新研究中发现在N末端乙酰转移酶的表达受到抑制后，线虫发育和繁殖都受到严重影响，这表明N末端乙酰化修饰在线虫发育和繁殖中发挥着重要作用。本课题拟结合线虫RNAi实验体系，采用我们创建的一种高效的N末端乙酰化修饰的组学分析方法(Zhang,2009;2010;2011;2013)，系统地鉴定线虫N末端乙酰化修饰的位点及程度，深入探讨不同N末端乙酰转移酶的底物特异性，并揭示其在发育繁殖中的调控机制。通过这一研究，将拓展我们对N末端乙酰化修饰功能的认识并为发育繁殖相关的研究提供新的思路。