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

Author

Listed:
  • Tatsuya Morisaki

    (Fluorescence Imaging Group, National Cancer Institute, NIH)

  • Waltraud G. Müller

    (Fluorescence Imaging Group, National Cancer Institute, NIH)

  • Nicole Golob

    (Institut für Pathologie Graz, 8036 Graz, Auenbruggerplatz 25/1, Austria)

  • Davide Mazza

    (Università Vita Salute San Raffaele e IRCCS Ospedale San Raffaele)

  • James G. McNally

    (Fluorescence Imaging Group, National Cancer Institute, NIH
    Present address: Institute for Soft Matter and Functional Materials, Helmholtz Center Berlin, Albert-Einstein-Str. 15, 12489 Berlin, Germany)

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.

Suggested Citation

  • Tatsuya Morisaki & Waltraud G. Müller & Nicole Golob & Davide Mazza & James G. McNally, 2014. "Single-molecule analysis of transcription factor binding at transcription sites in live cells," Nature Communications, Nature, vol. 5(1), pages 1-8, December.
    • Handle: RePEc:nat:natcom:v:5:y:2014:i:1:d:10.1038_ncomms5456
    DOI: 10.1038/ncomms5456
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    Cited by:

    1. Matteo Mazzocca & Alessia Loffreda & Emanuele Colombo & Tom Fillot & Daniela Gnani & Paola Falletta & Emanuele Monteleone & Serena Capozi & Edouard Bertrand & Gaelle Legube & Zeno Lavagnino & Carlo Ta, 2023. "Chromatin organization drives the search mechanism of nuclear factors," Nature Communications, Nature, vol. 14(1), pages 1-17, December.

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