Purification and characterisation of the Drosophila dMi2 chromatin remodelling machine

染色质结构的修饰对DNA转录、复制和修复的基本过程至关重要。染色质结构的改变涉及atp依赖的染色质重塑和组蛋白的翻译后修饰。然而，人们对其精确的分子机制和两者之间的关系知之甚少。该项目旨在分析Mi2重塑机的功能，Mi2重塑机是一种独特的蛋白质复合物，结合了atp依赖性重塑和组蛋白去乙酰化活性，并与细胞周期控制和肿瘤发生有关。三种方法将并行使用：(1)从果蝇胚胎提取物中分离含有dMi2重塑atp酶及其相关组蛋白去乙酰化酶的复合物，研究其亚基组成和活性。(2)在体外表达重组dMi2并对其活性进行表征。(3)果蝇细胞培养系统将用于研究dMi2和dRPD3在细胞周期控制中的作用。

The modification of chromatin structure is of paramount importance for the fundamental processes of DNA transcription, replication and repair. Alteration of chromatin structure involves ATP-dependent chromatin remodelling and posttranslational modification of histones. The precise molecular mechanism of and the relationship between the two is, however, poorly understood. This project aims to analyse the function of the Mi2 remodelling machine, a protein complex which is unique in combining both ATP-dependent remodelling and histone deacetylation activities and which has been implicated in cell cycle control and tumourigenesis. Three approaches will be employed in parallel: (1) Complexes containing the dMi2 remodelling ATPase and its associated histone deacetylase will be isolated from Drosophila embryo extracts to study their subunit composition and activities. (2) Recombinant dMi2 will be expressed and its activity will be characterized in vitro. (3) Drosophila cell culture systems will be used to investigate the role of dMi2 and dRPD3 in cell cycle control.