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Title: Biochemical analysis of MeCP2
Author: Klose, Robert John
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2005
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MeCP2 is a transcriptional repressor that recruits the Sin3a chromatin remodelling complex to methylated loci. Sin3a-associated histone deacetylases contribute to the ability of MeCP2 to repress transcription and modulate chromatin structure. The biomedical importance of normal MeCP2 function is highlighted by the discovery that inactivating mutations in MeCP2 cause the severe neurological disease Rett syndrome. By deleting the Mecp2 gene, a mouse model of Rett syndrome has been generated and used to study the molecular and physiological outcome of MeCP2 deficiency. Inefficient regulation of neuronal gene expression may have a role in the etiology of Rett syndrome. By studying the biochemical properties of MeCP2 this thesis addresses in detail three basic questions; (1) what are the native biochemical properties of MeCP2? (2) what specific DNA sequences does MeCP2 bind? and (3) what are the affects of post-translational modification on MeCP2? To address the composition of any mammalian MeCP2 complexes, native MeCP2 was purified to near homogeneity from rat brain. Native MeCP2 is an elongated monomer that does not stably associated with other cofactors including Sin3a. Analysis of MeCP2 binding sites in vivo demonstrates that MeCP2 binds unique loci when compared to other MBP’s. Using an unbiased in vitro DNA binding site evolution assay, Methyl-SELEX, MeCP2 was shown to require methyl-CpG sequences containing a flanking run of A/T rich DNA for high affinity binding. Finally, biochemical fractionation of nuclear proteins revealed activities that phosphorylate MeCP2, and the potential affects of this modification were explored.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available