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Series GSE31951 Query DataSets for GSE31951
Status Public on Sep 08, 2011
Title Genome-wide activity-dependent MeCP2 phosphorylation regulates nervous system development and function [ChIP-Seq]
Organism Mus musculus
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary Autism spectrum disorders such as Rett syndrome (RTT) have been hypothesized to arise from defects in experience-dependent synapse maturation. RTT is caused by mutations in MECP2, a nuclear protein that becomes phosphorylated at S421 in response to neuronal activation. We show here that disruption of MeCP2 S421 phosphorylation in vivo results in defects in synapse development and behavior, implicating activity-dependent regulation of MeCP2 in brain development and RTT. We investigated the mechanism by which S421 phosphorylation regulates MeCP2 function and show by chromatin immunoprecipitation-sequencing that this modification occurs on MeCP2 bound across the genome. The phosphorylation of MeCP2 S421 appears not to regulate the expression of specific genes; rather, MeCP2 functions as a histone-like factor whose phosphorylation may facilitate a genome-wide response of chromatin to neuronal activity during nervous system development. We propose that RTT results in part from a loss of this experience-dependent chromatin remodeling.
To examine MeCP2 binding across the neuronal genome and where on the genome MeCP2 is phosphorylated at Serine 421 in response to neuronal activity we performed anti-total MeCP2 and anti-phospho-Serine 421 specific Chromatin immunoprecipitation from cultured cortical neurons that were either left unstimulated or membrane depolarized for 2 hours by addition of 55mM KCl to the media. ChIP DNA was verified for successful IP by qPCR then cloned and sequenced using ABI SOLiD system 4.
 
Overall design ChIP was performed from E16 +7DIV disociated cortical cultures from one or two independent dissections using an anti-c-terminal antiserum recognizing MeCP2 independent of its phosphorylation state or with an anti-pS421 antiserum that specifically immunoprecipitates MeCP2 phosphorylated at serine 421. Samples were qPCR validated and sequenced using ABI SOLiD system 4 library preparation and sequencing.
 
Contributor(s) Cohen S, Hemberg M, Hutchinson AN, Sadacca LA, Ebert DH, Harmin DA, Greenberg RS, Zhou Z, Wetsel WC, West AE, Gabel HW, Greenberg ME
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Submission date Sep 07, 2011
Last update date May 15, 2019
Contact name Harrison Wren Gabel
E-mail(s) harrison_gabel@hms.harvard.edu
Phone 617-512-0289
Organization name Harvard Medical School
Department Neurobiology
Lab Michael Greenberg
Street address 7 Linden St Apt 3
City Brookline
State/province MA
ZIP/Postal code 02445
Country USA
 
Platforms (1)
GPL14602 AB SOLiD 4 System (Mus musculus)
Samples (9)
GSM791710 0hrKCl-Input-sampleB1
GSM791711 0hrKCl-MeCP2IP-sampleB1
GSM791712 2hrKcl-Input-sampleB1
This SubSeries is part of SuperSeries:
GSE31851 Genome-wide activity-dependent MeCP2 phosphorylation regulates nervous system development and function
Relations
SRA SRP008119
BioProject PRJNA155291

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE31951_RAW.tar 3.6 Gb (http)(custom) TAR (of BIGWIG)
SRA Run SelectorHelp
Processed data provided as supplementary file
Raw data are available in SRA

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