Purification and characterisation of the Drosophila dMi2 chromatin remodelling machine

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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.