Journal Article

DZNE-2024-01257

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png The Alpha-Synuclein Gene (SNCA) is a Genomic Target of Methyl-CpG Binding Protein 2 (MeCP2)-Implications for Parkinson's Disease and Rett Syndrome.

Schmitt, I. (First author)DZNE* ; Evert, B. O. ; Sharma, A. ; Khazneh, H. ; Murgatroyd, C. ; Wüllner, U. (Last author)DZNE*

2024
Humana Press Totowa, NJ

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Please use a persistent id in citations: doi:10.1007/s12035-024-03974-3

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.

Keyword(s): Methyl-CpG-Binding Protein 2: genetics (MeSH) ; Methyl-CpG-Binding Protein 2: metabolism (MeSH) ; alpha-Synuclein: metabolism (MeSH) ; alpha-Synuclein: genetics (MeSH) ; Humans (MeSH) ; Rett Syndrome: genetics (MeSH) ; Rett Syndrome: metabolism (MeSH) ; Cell Line, Tumor (MeSH) ; Parkinson Disease: genetics (MeSH) ; Parkinson Disease: metabolism (MeSH) ; DNA Methylation: genetics (MeSH) ; Protein Binding (MeSH) ; Introns: genetics (MeSH) ; Mutation: genetics (MeSH) ; SNCA ; Alpha-synuclein ; DNA methylation ; Epigenetic ; Genomic target ; Intron ; MeCP2 ; Methyl-CpG binding protein 2 ; Parkinson’s disease ; RTT ; Rett syndrome ; Methyl-CpG-Binding Protein 2 ; alpha-Synuclein ; SNCA protein, human ; MECP2 protein, human

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Contributing Institute(s):

1. Biomarker Parkinson's Disease (AG Wüllner)

Research Program(s):

1. 353 - Clinical and Health Care Research (POF4-353) (POF4-353)

Appears in the scientific report 2024

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Database coverage:
; ; ; BIOSIS Previews ; Biological Abstracts ; Clarivate Analytics Master Journal List ; DEAL Springer ; Essential Science Indicators ; IF >= 5 ; JCR ; SCOPUS ; Science Citation Index Expanded ; Web of Science Core Collection

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