Motif orientation matters: Structural characterization of TEAD1 recognition of genomic DNA
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
33333006
DOI
10.1016/j.str.2020.11.018
PII: S0969-2126(20)30465-2
Knihovny.cz E-resources
- Keywords
- DNA binding, M-CAT, TEAD1, chemical cross-linking and single molecule FRET, high- and low-affinity binding sites, hydrogen-deuterium exchange, native electrospray, structural mass spectrometry, transcription factor,
- MeSH
- DNA-Binding Proteins chemistry metabolism MeSH
- DNA chemistry metabolism MeSH
- Nuclear Proteins chemistry metabolism MeSH
- Jurkat Cells MeSH
- Humans MeSH
- Nucleotide Motifs MeSH
- Molecular Docking Simulation MeSH
- TEA Domain Transcription Factors MeSH
- Transcription Factors chemistry metabolism MeSH
- Protein Binding MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- DNA-Binding Proteins MeSH
- DNA MeSH
- Nuclear Proteins MeSH
- TEAD1 protein, human MeSH Browser
- TEA Domain Transcription Factors MeSH
- Transcription Factors MeSH
TEAD transcription factors regulate gene expression through interactions with DNA and other proteins. They are crucial for the development of eukaryotic organisms and to control the expression of genes involved mostly in cell proliferation and differentiation; however, their deregulation can lead to tumorigenesis. To study the interactions of TEAD1 with M-CAT motifs and their inverted versions, the KD of each complex was determined, and H/D exchange, quantitative chemical cross-linking, molecular docking, and smFRET were utilized for structural characterization. ChIP-qPCR was employed to correlate the results with a cell line model. The results obtained showed that although the inverted motif has 10× higher KD, the same residues were affected by the presence of M-CAT in both orientations. Molecular docking and smFRET revealed that TEAD1 binds the inverted motif rotated 180°. In addition, the inverted motif was proven to be occupied by TEAD1 in Jurkat cells, suggesting that the low-affinity binding sites present in the human genome may possess biological relevance.
Institute of Biotechnology Czech Academy of Sciences Vestec 252 50 Czech Republic
Institute of Microbiology Czech Academy of Sciences Prague 142 20 Czech Republic
J Heyrovsky Institute of Physical Chemistry Czech Academy of Sciences Prague 182 00 Czech Republic
University of Connecticut Department of Chemistry 55 N Eagleville Road Storrs CT 06269 USA
References provided by Crossref.org
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