Motif orientation matters: Structural characterization of TEAD1 recognition of genomic DNA - Structural MS Supporting Data

Supporting data for the paper "Motif orientation matters: Structural characterization of TEAD1 recognition of genomic DNA" (Filandrova et al.).

Summary:
TEAD transcription factors regulate gene expression through interactions with DNA and other proteins. They are crucial for development of eukaryotic organisms and control expression of genes involved mostly in cell proliferation and differentiation, however their deregulation can lead to tumorigenesis. To study the interaction of TEAD1 with M-CAT motifs and their inverted versions 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 cell line model. Obtained results showed that although the inverted motif has 10× higher KD, the same residues were affected by presence of M-CAT in both orientations. Molecular docking and smFRET revealed that TEAD1 binds the inverted motif 180° rotated. Additionally, the inverted motif was proved to be occupied by TEAD1 in Jurkat cells, suggesting that the low-affinity binding sites present in human genome may possess biological relevance.

Here, we deposit supporting data for the structural mass spectrometry experiments containing:

1) Mass spectra of cross-links dissociation products. Each spectrum represents a single identified cross-link consisting of two peptides connected with the disuccinimidyl adipate (DSA) reagent. Fragmentation sites are marked along the amino acids sequence of each peptide. Resulting fragments are highlighted in the spectrum. (Figure MD-1, page 1)

2) Deuterium uptake plots of all identified peptides. Comparison of unbound state and complexes of TEAD1-DBD with different M-CATs. Regions, where the curves are not aligned, show decreased solvent accessibility in presence of DNA. NT stands for short N-terminal region arising from thrombin cleavage site. Deviations between triplicates are displayed as error bars. (Figure MD-2, page 9)