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@ARTICLE{Schmitt:272839,
author = {Schmitt, Ina and Evert, Bernd O and Sharma, Amit and
Khazneh, Hassan and Murgatroyd, Chris and Wüllner, Ullrich},
title = {{T}he {A}lpha-{S}ynuclein {G}ene ({SNCA}) is a {G}enomic
{T}arget of {M}ethyl-{C}p{G} {B}inding {P}rotein 2
({M}e{CP}2)-{I}mplications for {P}arkinson's {D}isease and
{R}ett {S}yndrome.},
journal = {Molecular neurobiology},
volume = {61},
number = {10},
issn = {0893-7648},
address = {Totowa, NJ},
publisher = {Humana Press},
reportid = {DZNE-2024-01257},
pages = {7830 - 7844},
year = {2024},
abstract = {Mounting evidence suggests a prominent role for
alpha-synuclein (a-syn) in neuronal cell function.
Alterations in the levels of cellular a-syn have been
hypothesized to play a critical role in the development of
Parkinson's disease (PD); however, mechanisms that control
expression of the gene for a-syn (SNCA) in cis and trans as
well as turnover of a-syn are not well understood. We
analyzed whether methyl-CpG binding protein 2 (MeCP2), a
protein that specifically binds methylated DNA, thus
regulating transcription, binds at predicted binding sites
in intron 1 of the SNCA gene and regulates a-syn protein
expression. Chromatin immunoprecipitation (ChIP) and
electrophoretic mobility-shift assays (EMSA) were used to
confirm binding of MeCP2 to regulatory regions of SNCA.
Site-specific methylation and introduction of localized
mutations by CRISPR/Cas9 were used to investigate the
binding properties of MeCP2 in human SK-N-SH neuroblastoma
cells. The significance of MeCP2 for SNCA regulation was
further investigated by overexpressing MeCP2 and mutated
variants of MeCP2 in MeCP2 knockout cells. We found that
methylation-dependent binding of MeCP2 at a restricted
region of intron 1 of SNCA had a significant impact on the
production of a-syn. A single nucleotide substitution near
to CpG1 strongly increased the binding of MeCP2 to intron 1
of SNCA and decreased a-syn protein expression by $60\%.$ In
contrast, deletion of a single nucleotide closed to CpG2 led
to reduced binding of MeCP2 and significantly increased
a-syn levels. In accordance, knockout of MeCP2 in SK-N-SH
cells resulted in a significant increase in a-syn
production, demonstrating that SNCA is a genomic target for
MeCP2 regulation. In addition, the expression of two mutated
MeCP2 variants found in Rett syndrome (RTT) showed a loss of
their ability to reduce a-syn expression. This study
demonstrates that methylation of CpGs and binding of MeCP2
to intron 1 of the SNCA gene plays an important role in the
control of a-syn expression. In addition, the changes in
SNCA regulation found by expression of MeCP2 variants
carrying mutations found in RTT patients may be of
importance for the elucidation of a new molecular pathway in
RTT, a rare neurological disorder caused by mutations in
MECP2.},
keywords = {Methyl-CpG-Binding Protein 2: genetics / Methyl-CpG-Binding
Protein 2: metabolism / alpha-Synuclein: metabolism /
alpha-Synuclein: genetics / Humans / Rett Syndrome: genetics
/ Rett Syndrome: metabolism / Cell Line, Tumor / Parkinson
Disease: genetics / Parkinson Disease: metabolism / DNA
Methylation: genetics / Protein Binding / Introns: genetics
/ Mutation: genetics / SNCA (Other) / Alpha-synuclein
(Other) / DNA methylation (Other) / Epigenetic (Other) /
Genomic target (Other) / Intron (Other) / MeCP2 (Other) /
Methyl-CpG binding protein 2 (Other) / Parkinson’s disease
(Other) / RTT (Other) / Rett syndrome (Other) /
Methyl-CpG-Binding Protein 2 (NLM Chemicals) /
alpha-Synuclein (NLM Chemicals) / SNCA protein, human (NLM
Chemicals) / MECP2 protein, human (NLM Chemicals)},
cin = {AG Wüllner},
ddc = {570},
cid = {I:(DE-2719)1011302},
pnm = {353 - Clinical and Health Care Research (POF4-353)},
pid = {G:(DE-HGF)POF4-353},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:38429622},
pmc = {pmc:PMC11415397},
doi = {10.1007/s12035-024-03974-3},
url = {https://pub.dzne.de/record/272839},
}