Brain phosphorylation of MeCP2 at serine 164 is developmentally regulated and globally alters its chromatin association

Stefanelli, Gilda; Gandaglia, Anna; Costa, Mario; Cheema, Manjinder S.; Marino, Daniele Di; Barbiero, Isabella; Kilstrup‐Nielsen, Charlotte; Ausió, Juan; Landsberger, Nicoletta

doi:10.1038/srep28295

Cited by 26 publications

(24 citation statements)

References 47 publications

(84 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3 became MeCP2 phosphorylation. [65][66][67] MeCP2 is an intrinsically disordered protein and, as such, is subject to many PTMs, 7 including phosphorylation. 67 Histone phosphorylation has been shown to globally decrease its chromatin binding affinity 68,69 and TSA has been shown to downregulate several cyclin dependent kinases (CDKs).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Trichostatin A decreases the levels of MeCP2 expression and phosphorylation and increases its chromatin binding affinity

et al. 2017

Self Cite

View full text Add to dashboard Cite

MeCP2 binds to methylated DNA in a chromatin context and has an important role in cancer and brain development and function. Histone deacetylase (HDAC) inhibitors are currently being used to palliate many cancer and neurological disorders. Yet, the molecular mechanisms involved are not well known for the most part and, in particular, the relationship between histone acetylation and MeCP2 is not well understood. In this paper, we study the effect of the HDAC inhibitor trichostatin A (TSA) on MeCP2, a protein whose dysregulation plays an important role in these diseases. We find that treatment of cells with TSA decreases the phosphorylation state of this protein and appears to result in a higher MeCP2 chromatin binding affinity. Yet, the binding dynamics with which the protein binds to DNA appear not to be significantly affected despite the chromatin reorganization resulting from the high levels of acetylation. HDAC inhibition also results in an overall decrease in MeCP2 levels of different cell lines. Moreover, we show that miR132 increases upon TSA treatment, and is one of the players involved in the observed downregulation of MeCP2.

show abstract

Section: Resultsmentioning

confidence: 99%

“…69 We also decided to include S164-P, a recently described abundant developmental MeCP2 phosphorylation, which has been documented to lower its binding affinity to chromatin. 66 The results of such analyses are shown in Fig. 4.…”

Section: Resultsmentioning

confidence: 99%

Trichostatin A decreases the levels of MeCP2 expression and phosphorylation and increases its chromatin binding affinity

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Mecp2 Post-translational Modificationsmentioning

confidence: 99%

“…A conserved serine (S164) located at the beginning of ID just after the MBD, was shown to be abundantly phosphorylated in the brain in a developmentally regulated manner [97]. While the phospho-mimicking version S164D showed minor binding to chromatin in live-cell kinetic studies, the phospho-defective mutation S164A had the opposite effect [97]. These results could be explained by in silico modeling of the 3D structure of this phosphorylation site, revealing the addition of negative charge to the protein surface as a consequence of S164 phosphorylation, hence, decreasing DNA binding.…”

Section: Mecp2 Post-translational Modificationsmentioning

confidence: 99%

“…These results could be explained by in silico modeling of the 3D structure of this phosphorylation site, revealing the addition of negative charge to the protein surface as a consequence of S164 phosphorylation, hence, decreasing DNA binding. Immunofluorescence analysis of wildtype neurons versus MeCP2 S164 mutants revealed that temporal regulation of S164 phosphorylation is required for proper nuclear size and neuronal dendritic branching [97].…”

Section: Mecp2 Post-translational Modificationsmentioning

confidence: 99%

See 1 more Smart Citation

MeCP2 and Chromatin Compartmentalization

Schmidt

Zhang

Cardoso

2020

Cells

View full text Add to dashboard Cite

Methyl-CpG binding protein 2 (MeCP2) is a multifunctional epigenetic reader playing a role in transcriptional regulation and chromatin structure, which was linked to Rett syndrome in humans. Here, we focus on its isoforms and functional domains, interactions, modifications and mutations found in Rett patients. Finally, we address how these properties regulate and mediate the ability of MeCP2 to orchestrate chromatin compartmentalization and higher order genome architecture.Cells 2020, 9, 878 2 of 31 the predominant isoform in brain and has an earlier expression onset than MeCP2 e2 [6]. The two isoforms are commonly considered as functionally equivalent, yet recent evidence shows that MeCP2 e1 plays a role in neuronal maturation [7] and is more relevant for RTT [8][9][10]. In view of the fact that MeCP2 e2 isoform was the first to be known and a much larger body of literature pertains to this isoform, we will, throughout, use amino acid coordinates from MeCP2 e2 isoform.Both variants include two functionally characterized domains: the methyl-CpG binding domain (MBD) and the transcriptional repression domain (TRD). The MBD specifically recognizes and binds 5-methylcytosine (5mC), while the TRD was found to bind multiple transcriptional repressors, thus silencing gene expression [11][12][13][14][15][16]. However, the TRD was also shown to bind to multiple transcriptional activators and activate gene expression [17][18][19]. More recently, the TRD has been narrowed down to the N-CoR/SMRT interacting domain (NID) [20]. A summary of the best characterized domains of MeCP2 is shown in Figure 1. The DNA binding properties of the different domains and the mechanism of DNA binding will be addressed in the next section. MeCP2 DNA DindingEarly studies on MeCP2 characterized it as a protein being capable to bind to a single, symmetrically methylated CpG pair via the MBD domain spanning amino acids 89 -162 and thereby overlapping approximately twelve base pairs of DNA [4,21,22]. Later studies indicated that the N-terminal domain (NTD) enhanced DNA binding affinity via the MBD [23], while the intervening domain (ID), TRD and C-terminal domain (CTD) alpha showed methylation-independent DNA binding capabilities and CTD beta was proposed to bind to chromatin, but not to naked DNA [23,24]. Furthermore, three AT-hook-like domains were identified within the ID, TRD and CTD alpha domains (AT-hook 1, aa 184-195; AT-hook 2, aa 264-273; AT-hook 3, aa 341-364). The AT-hook motif is a short motif binding to the minor groove of AT-rich DNA via the core consensus amino acid sequence RGRP [25]. These methylation-independent DNA binding capabilities allow MeCP2 to bind to different sites on the DNA at the same time, thus, possibly contributing to genome-wide chromatin organization. With the exception of the MBD, MeCP2 was shown to be mostly an intrinsically disordered protein. Upon binding to DNA, though, increased secondary structure in ID and TRD were observed [23]. The MBD is the only domain showing structurally conserved motifs, as it cont...

show abstract