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Nucleic Acids Res ; 48(15): 8529-8544, 2020 09 04.
Artículo en Inglés | MEDLINE | ID: mdl-32738045

RESUMEN

Myocyte enhancer factor-2B (MEF2B) has the unique capability of binding to its DNA target sites with a degenerate motif, while still functioning as a gene-specific transcriptional regulator. Identifying its DNA targets is crucial given regulatory roles exerted by members of the MEF2 family and MEF2B's involvement in B-cell lymphoma. Analyzing structural data and SELEX-seq experimental results, we deduced the DNA sequence and shape determinants of MEF2B target sites on a high-throughput basis in vitro for wild-type and mutant proteins. Quantitative modeling of MEF2B binding affinities and computational simulations exposed the DNA readout mechanisms of MEF2B. The resulting binding signature of MEF2B revealed distinct intricacies of DNA recognition compared to other transcription factors. MEF2B uses base readout at its half-sites combined with shape readout at the center of its degenerate motif, where A-tract polarity dictates nuances of binding. The predominant role of shape readout at the center of the core motif, with most contacts formed in the minor groove, differs from previously observed protein-DNA readout modes. MEF2B, therefore, represents a unique protein for studies of the role of DNA shape in achieving binding specificity. MEF2B-DNA recognition mechanisms are likely representative for other members of the MEF2 family.


Asunto(s)
Proteínas de Unión al ADN/ultraestructura , ADN/ultraestructura , Complejos Multiproteicos/ultraestructura , Secuencia de Aminoácidos/genética , Sitios de Unión/genética , ADN/genética , Proteínas de Unión al ADN/química , Humanos , Linfoma de Células B/genética , Linfoma de Células B/patología , Proteínas de Dominio MADS/genética , Proteínas de Dominio MADS/ultraestructura , Factores de Transcripción MEF2/química , Factores de Transcripción MEF2/ultraestructura , Complejos Multiproteicos/genética , Conformación de Ácido Nucleico , Motivos de Nucleótidos/genética , Unión Proteica/genética
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