TY - JOUR
T1 - DNA residence time is a regulatory factor of transcription repression
AU - Clauß, Karen
AU - Popp, Achim P.
AU - Schulze, Lena
AU - Hettich, Johannes
AU - Reisser, Matthias
AU - Torres, Laura Escoter
AU - Uhlenhaut, N. Henriette
AU - Gebhardt, J. Christof M.
N1 - Publisher Copyright:
© The Author(s) 2017.
PY - 2017/11/2
Y1 - 2017/11/2
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85038028954&partnerID=8YFLogxK
U2 - 10.1093/nar/gkx728
DO - 10.1093/nar/gkx728
M3 - Article
C2 - 28977492
AN - SCOPUS:85038028954
SN - 0305-1048
VL - 45
SP - 11121
EP - 11130
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 19
ER -