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Nat Commun. 2014 Jul 18;5:4456. doi: 10.1038/ncomms5456.

Single-molecule analysis of transcription factor binding at transcription sites in live cells.

Author information

1
1] Fluorescence Imaging Group, National Cancer Institute, NIH, Bethesda, Maryland 20892, USA [2].
2
Institut für Pathologie Graz, 8036 Graz, Auenbruggerplatz 25/1, Austria.
3
Università Vita Salute San Raffaele e IRCCS Ospedale San Raffaele, 20132 Milano, Italy.

Abstract

Although numerous live-cell measurements have shown that transcription factors (TFs) bind chromatin transiently, no measurements of transient binding have been reported at the endogenous response elements (REs) where transcription is normally induced. Here we show that at endogenous REs the transcriptionally productive specific binding of two TFs, p53 and the glucocorticoid receptor (GR), is transient. We also find that the transient residence times of GR at endogenous REs are roughly comparable to those at an artificial, multi-copy array of gene regulatory sites, supporting the use of multi-copy arrays for live-cell analysis of transcription. Finally, we find that at any moment only a small fraction of TF molecules are engaged in transcriptionally productive binding at endogenous REs. The small fraction of bound factors provides one explanation for gene bursting and it also indicates that REs may often be unoccupied, resulting in partial responses to transcriptional signals.

PMID:
25034201
PMCID:
PMC4144071
DOI:
10.1038/ncomms5456
[Indexed for MEDLINE]
Free PMC Article
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