Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.596825
Title: MeCP2 and its interaction with Sin3a, chromatin and DNA
Author: Bowers, Sarion Rebecca
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2007
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Abstract:
The ability of MeCP2 to repress transcription is in part due to its interaction with mSin3a – a transcriptional co-repressor with multiple binding partners, including histone deacetylases, which promote a compact, repressive chromatin structure. The Sin3a-interacting domain of MeCP2 is the transcription repression domain (TRD) but the region(s) through which Sin3a interacts with MeCp2 is (are) so far poorly – defined. In Chapter 2 experiments aimed at further defining this interaction are described. Two regions of mSin3a (amino acids 524-694 and 1002-1219) were found to interact with the TRD. Two regions of approximately 20 amino acids were involved in the binding of mSin3a to MeCP2 (amino acids 634-654 and 1041-1081). In Chapter 3, the possibility that MeCP2 is able to displace histone H1 from native and reconstituted chromatin was studied. The ability of MeCP2 to displace H1 from mono- and poly-nucleosomes (6-10 nucleosomes) was investigated using a sucrose gradient sedimentation assay. These assays and gel shift assays using nucleosomal arrays reconstituted on to a DNA template containing 200 bp repeats of the ‘601’ nucleosome positioning sequence, suggest that MeCP2 is partially able to displace H1. The MBD of MeCP2 is known to interact with four-way junction DNA in a methylation-independent structure-specific manner. Chapter 4 describes the use of 2D 15N HSQC spectra and chemical-shift perturbation analysis to investigate the interaction of MBD with four-way junction DNA. A region of the C-terminal half of the MBD, and the region previously identified as interacting with duplex DNA, were both found to be involved in the interaction with the four-way junction.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.596825  DOI: Not available
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