Cell Cycle proteomicsin Xenopus

描述（由申请人提供）： 蛋白质组学对非洲爪蟾研究具有特殊意义，因为减数分裂、受精和早期发育中的关键事件是在翻译、翻译后修饰和蛋白质水解水平上调控的，而不是通过转录。蛋白质组学是系统地研究这些水平的调节和发现新机制的唯一工具。我们建议通过在人类和酵母系统中开发的迁移技术，为非洲爪蟾胚胎和卵提取物开发一个尖端的蛋白质组学研究平台，并公开传播我们开发的数据库和工具。我们还将优化鸡蛋提取物中蛋白质复合物和隔室的蛋白质组学分析方法。使用这些工具，我们将在正常和扰动条件下定量早期发育和卵提取物中> 5，000个蛋白质，> 10，000个磷酸化位点和> 1，500个泛素化位点的相对量。我们将研究细胞周期调控的机制，细胞周期在早期发育过程中如何变化，微管如何成核，以及胚胎如何检测细胞核与细胞质比例的变化。

DESCRIPTION (provided by applicant): Proteomics has special significance for Xenopus research because key events in meiosis, fertilization and early development are regulated at the level of translation, post- translational modification and proteolysis, and not by transcription. Proteomics is the only tool for systematically investigating these levels of regulation and discovering new mechanisms. We propose to develop a cutting edge proteomic research platform for Xenopus embryos and egg extract by migrating technology developed in human and yeast systems, and to publicly disseminate the databases and tools we develop. We will also optimize methods for proteomic analysis of protein complexes and compartments in egg extract. Using these tools we will quantify relative amounts of >5,000 proteins, >10,000 phosphorylation sites and >1,500 ubiquitination sites in early development and in egg extracts under normal and perturbed conditions. We will investigate the mechanism of cell cycle regulation, how the cell cycle changes during early development, how microtubules are nucleated, and how embryos detect changes in the nucleus to cytoplasm ratio.