Purification and characterisation of the Drosophila dMi2 chromatin remodelling machine

果蝇 dMi2 染色质重塑机的纯化和表征

• 批准号: 5275356
• 负责人: Professor Dr. Alexander Brehm
• 金额: --
• 依托单位: Institut für Molekularbiologie und Tumorforschung (IMT)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2000
• 资助国家: 德国
• 起止时间: 1999-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5275356?language=en
• 关键词: Purification; characterisation; Drosophila; dMi2; chromatin

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.

染色质结构的修饰对于 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 依赖性染色质重塑和组蛋白翻译后修饰。然而，人们对两者之间的精确分子机制和关系知之甚少。该项目旨在分析 Mi2 重塑机器的功能，Mi2 重塑机器是一种蛋白质复合物，独特地将 ATP 依赖性重塑和组蛋白脱乙酰化活性结合在一起，并且与细胞周期控制和肿瘤发生有关。将同时采用三种方法：（1）将从果蝇胚胎提取物中分离出含有 dMi2 重塑 ATP 酶及其相关组蛋白脱乙酰酶的复合物，以研究其亚基组成和活性。 (2) 重组dMi2将在体外表达并表征其活性。 (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。