Cell Cycle proteomicsin Xenopus

描述(由申请人提供)： 蛋白质组学对非洲爪哇的研究具有特殊的意义，因为非洲爪哇减数分裂、受精和早期发育中的关键事件是在翻译、翻译后修饰和蛋白分解水平上调节的，而不是通过转录调节的。蛋白质组学是系统研究这些调控水平和发现新机制的唯一工具。我们建议通过移植人类和酵母系统中开发的技术，为非洲爪哇胚胎和鸡蛋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体和隔间的蛋白质组学分析方法。使用这些工具，我们将量化&gt；5,000个蛋白质、&gt；10,000个磷酸化位点和&gt；1,500个泛素化位点在发育早期以及在正常和扰动条件下的鸡蛋提取物中的相对数量。我们将研究细胞周期调节的机制，细胞周期在早期发育过程中如何变化，微管是如何成核的，以及胚胎如何检测核质比的变化。 公共卫生相关性： 项目简介我们将开发基础广泛的工具，用于研究细胞分裂周期中蛋白质水平的变化，将革命性的质谱学技术引入胚胎系统。这类研究可以为设计影响癌症和炎症重要途径的药物提供更好的基础。

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. PUBLIC HEALTH RELEVANCE: Project Narrative We will develop broad based tools for the study of changes in the protein levels during the cell division cycle, bringing the revolutionary technique of mass spectrometry to embryonic systems. Such studies could offer a better basis for designing drugs that affect important pathways in cancer and inflammation.