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 K_D 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 K_D, 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.

TEAD 转录因子通过与 DNA 和其他蛋白质的相互作用来调节基因表达。它们对于真核生物的发育和控制主要参与细胞增殖和分化的基因的表达至关重要；然而，它们的失调会导致肿瘤发生。为了研究 TEAD1 与 M-CAT 基序及其反向版本的相互作用，确定了每个复合物的 K _D，并利用 H/D 交换、定量化学交联、分子对接和 smFRET 进行结构表征。ChIP-qPCR 用于将结果与细胞系模型相关联。获得的结果表明，虽然倒置基序的 K _D高出 10 倍，相同的残基在两个方向都受到 M-CAT 的影响。分子对接和 smFRET 显示 TEAD1 结合旋转 180° 的倒置基序。此外，倒置基序被证明在 Jurkat 细胞中被 TEAD1 占据，这表明人类基因组中存在的低亲和力结合位点可能具有生物学相关性。