NRSA Fellowship for Reidmann: Determination of the epigenetic regulation of gene transcription by MeCP2 in neurons

The most common neurodevelopment disorder in girls, Rett Syndrome (RTT), is caused by de novo loss of function mutations in methyl CpG binding protein 2 (MeCP2), which is found in high abundance in fully differentiated neurons. While the molecular mechanism of MeCP2 remains unknown widespread transcriptional deregulation occurs in neurons with mutated MeCP2. As a chromatin architectural protein (CAP) MeCP2s bind the DNA entry/exit dyad of the nucleosome and has the ability to regulate nucleosome stability and higher order chromatin structures. We hypothesize that MeCP2 regulates nucleosome breathing, stability, and composition to create and maintain specific chromatin structures critical in proper transcriptional regulation. We propose a variety of studies, including genome wide nucleosome positioning assays, FRET, AUC, electromobility gel shift, and restriction enzyme digests, to study the effect of MeCP2 on the individual nucleosome, local chromatin structure, and global chromatin. Together our studies will provide detailed information on the genome-wide impact of MeCP2, and further determine the mechanism by which MeCP2 regulates transcription.