Data from: Effective In vivo binding energy landscape illustrates kinetic stability of RBPJ-DNA binding

Transcription factors (TFs) such as RBPJ in Notch signaling bind to
specific DNA sequences to regulate transcription. How TF-DNA binding
kinetics and cofactor interactions modulate gene regulation is mostly
unknown. We determined the binding kinetics, transcriptional activity, and
genome-wide chromatin occupation of RBPJ and mutant variants by live-cell
single-molecule tracking, reporter assays, and ChIP-Seq. Importantly, the
search time of RBPJ exceeded its residence time, indicating kinetic rather
than thermodynamic binding stability. Impaired RBPJ-DNA binding as in
Adams-Oliver-Syndrome affected both target site association and
dissociation, while impaired cofactor binding mainly altered association
and unspecific binding. Moreover, our data point to the possibility that
cofactor binding contributes to target site specificity. Findings for
other TFs comparable to RBPJ indicate that kinetic rather than
thermodynamic DNA binding stability might prevail in vivo. We propose an
effective in vivo binding energy landscape of TF-DNA interactions as
instructive visualization of binding kinetics and mutation-induced
changes.