PuSH - Publikationsserver des Helmholtz Zentrums München: DNA residence time is a regulatory factor of transcription repression.

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Clauß, K.* ; Popp, A.P.* ; Schulze, L.* ; Hettich, J.* ; Reisser, M.* ; Escoter Torres, L. ; Uhlenhaut, N.H. ; Gebhardt, J.C.M.*

DNA residence time is a regulatory factor of transcription repression.

Nucleic Acids Res. 45, 11121-11130 (2017)

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Transcription comprises a highly regulated sequence of intrinsically stochastic processes, resulting in bursts of transcription intermitted by quiescence. In transcription activation or repression, a transcription factor binds dynamically to DNA, with a residence time unique to each factor. Whether the DNA residence time is important in the transcription process is unclear. Here, we designed a series of transcription repressors differing in their DNA residence time by utilizing the modular DNA binding domain of transcription activator-like effectors (TALEs) and varying the number of nucleotide-recognizing repeat domains. We characterized the DNA residence times of our repressors in living cells using single molecule tracking. The residence times depended non-linearly on the number of repeat domains and differed by more than a factor of six. The factors provoked a residence time-dependent decrease in transcript level of the glucocorticoid receptor-activated gene SGK1. Down regulation of transcription was due to a lower burst frequency in the presence of long binding repressors and is in accordance with a model of competitive inhibition of endogenous activator binding. Our single molecule experiments reveal transcription factor DNA residence time as a regulatory factor controlling transcription repression and establish TALE-DNA binding domains as tools for the temporal dissection of transcription regulation.

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