A screen for MeCP2-TBL1 interaction inhibitors using a luminescence-based assay.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
08 03 2023
08 03 2023
Historique:
received:
16
11
2022
accepted:
13
02
2023
entrez:
8
3
2023
pubmed:
9
3
2023
medline:
11
3
2023
Statut:
epublish
Résumé
Understanding the molecular pathology of neurodevelopmental disorders should aid the development of therapies for these conditions. In MeCP2 duplication syndrome (MDS)-a severe autism spectrum disorder-neuronal dysfunction is caused by increased levels of MeCP2. MeCP2 is a nuclear protein that binds to methylated DNA and recruits the nuclear co-repressor (NCoR) complex to chromatin via an interaction with the WD repeat-containing proteins TBL1 and TBLR1. The peptide motif in MeCP2 that binds to TBL1/TBLR1 is essential for the toxicity of excess MeCP2 in animal models of MDS, suggesting that small molecules capable of disrupting this interaction might be useful therapeutically. To facilitate the search for such compounds, we devised a simple and scalable NanoLuc luciferase complementation assay for measuring the interaction of MeCP2 with TBL1/TBLR1. The assay allowed excellent separation between positive and negative controls, and had low signal variance (Z-factor = 0.85). We interrogated compound libraries using this assay in combination with a counter-screen based on luciferase complementation by the two subunits of protein kinase A (PKA). Using this dual screening approach, we identified candidate inhibitors of the interaction between MeCP2 and TBL1/TBLR1. This work demonstrates the feasibility of future screens of large compound collections, which we anticipate will enable the development of small molecule therapeutics to ameliorate MDS.
Identifiants
pubmed: 36890145
doi: 10.1038/s41598-023-29915-z
pii: 10.1038/s41598-023-29915-z
pmc: PMC9995496
doi:
Substances chimiques
Receptors, Cytoplasmic and Nuclear
0
Repressor Proteins
0
Methyl-CpG-Binding Protein 2
0
Nuclear Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3868Subventions
Organisme : Wellcome Trust
ID : Centre grant (091580/Z/10/Z)
Pays : United Kingdom
Organisme : European Research Council
ID : EC 694295 Gen-Epix
Pays : International
Organisme : Wellcome Trust
ID : 107930/Z/15/ Z
Pays : United Kingdom
Informations de copyright
© 2023. The Author(s).
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