Brain phosphorylation of MeCP2 at serine 164 is developmentally regulated and globally alters its chromatin association.
MeCP2 is a transcriptional regulator whose functional alterations are responsible for several autism spectrum and mental disorders. Post-translational modifications (PTMs), and particularly differential phosphorylation, modulate MeCP2 function in response to diverse stimuli. Understanding the detailed role of MeCP2 phosphorylation is thus instrumental to ascertain how MeCP2 integrates the environmental signals and directs its adaptive transcriptional responses. The evolutionarily conserved serine 164 (S164) was found phosphorylated in rodent brain but its functional role has remained uncharacterized. We show here that phosphorylation of S164 in brain is dynamically regulated during neuronal maturation. S164 phosphorylation highly impairs MeCP2 binding to DNA in vitro and largely affects its nucleosome binding and chromatin affinity in vivo. Strikingly, the chromatin-binding properties of the global MeCP2 appear also extensively altered during the course of brain maturation. Functional assays reveal that proper temporal regulation of S164 phosphorylation controls the ability of MeCP2 to regulate neuronal morphology. Altogether, our results support the hypothesis of a complex PTM-mediated functional regulation of MeCP2 potentially involving a still poorly characterized epigenetic code. Furthermore, they demonstrate the relevance of the Intervening Domain of MeCP2 for binding to DNA.
San Raffaele Rett Research Unit, San Raffaele Scientific Institute, Milan, Italy.
Gilda
G
Stefanelli
San Raffaele Rett Research Unit, San Raffaele Scientific Institute, Milan, Italy.
Anna
A
Gandaglia
Institute of Neuroscience, National Research Council (CNR), Scuola Normale Superiore Pisa, Italy.
Mario
M
Costa
Department of Biochemistry and Microbiology, University of Victoria, Victoria (BC), Canada.
Manjinder S
MS
Cheema
Department of Informatics, Institute of Computational Science, Università della Svizzera Italiana, Lugano, Switzerland.
Daniele
D
Di Marino
Department of Biotechnology and Life Sciences, University of Insubria, Busto Arsizio (VA), Italy.
Isabella
I
Barbiero
Department of Biotechnology and Life Sciences, University of Insubria, Busto Arsizio (VA), Italy.
Charlotte
C
Kilstrup-Nielsen
Department of Biochemistry and Microbiology, University of Victoria, Victoria (BC), Canada.
Juan
J
Ausió
San Raffaele Rett Research Unit, San Raffaele Scientific Institute, Milan, Italy.|Department of Medical Biotechnology and Translational Medicine, University of Milan, Milan, Italy.
Nicoletta
N
Landsberger
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D019175:DNA Methylation; D003712:Dendrites / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2 / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D041681:NIH 3T3 Cells; D009707:Nucleosomes / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D011817:Rabbits; D012694:Serine / Q000378:metabolism
D002843:Chromatin; C497393:Mecp2 protein, mouse; D051783:Methyl-CpG-Binding Protein 2; D009707:Nucleosomes; D012694:Serine