CaltechAUTHORS
Published June 12, 2014 | Version Supplemental Material + Published
Journal Article Open

Single-molecule tracking in live cells reveals distinct target-search strategies of transcription factors in the nucleus

Creators

  • 1. ROR icon Institut de Biologie de l'École Normale Supérieure
  • 2. ROR icon Kastler-Brossel Laboratory
  • 3. ROR icon Janelia Research Campus
  • 4. ROR icon University of Paris
  • 5. ROR icon Laboratory of Theoretical Physics of Condensed Matter
  • 6. ROR icon Institute Curie

Abstract

Gene regulation relies on transcription factors (TFs) exploring the nucleus searching their targets. So far, most studies have focused on how fast TFs diffuse, underestimating the role of nuclear architecture. We implemented a single-molecule tracking assay to determine TFs dynamics. We found that c-Myc is a global explorer of the nucleus. In contrast, the positive transcription elongation factor P-TEFb is a local explorer that oversamples its environment. Consequently, each c-Myc molecule is equally available for all nuclear sites while P-TEFb reaches its targets in a position-dependent manner. Our observations are consistent with a model in which the exploration geometry of TFs is restrained by their interactions with nuclear structures and not by exclusion. The geometry-controlled kinetics of TFs target-search illustrates the influence of nuclear architecture on gene regulation, and has strong implications on how proteins react in the nucleus and how their function can be regulated in space and time.

Additional Information

© 2014, Izeddin et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. Received: January 7, 2014. Accepted: June 11, 2014. Accepted Manuscript published: June 12, 2014 (version 1). Version of Record published: July 15, 2014 (version 2). We would like to thank Dan Larson, John Lis, Florian Mueller, Yitzhak Rabin, Robert Singer, and Robert Tjian for discussions and comments. Leonid Mirny for an in depth review and his challenge and help with the modeling approach. We are grateful to Sarah Moorehead and Mohamed El Beheiry for critical reading and to Daniel Ciepielewski and Philippe Rideau for discussions on the microscopy. VR acknowledges financial support from FRM, and II from NWO. Work presented here was supported by the PCV DYNAFT 08-PCVI-0013 and DynamIC ANR-12-BSV5-0018 from Agence Nationale pour la Recherche, and a research contract with Nikon France to XD and MD. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Author contributions. II, VR, Conception and design, Acquisition of data, Analysis and interpretation of data, Drafting or revising the article; LB, CD-D, Acquisition of data, Drafting or revising the article; IIC, OB, RV, Analysis and interpretation of data, Drafting or revising the article; LB, Acquisition of data, Analysis and interpretation of data; FP, Acquisition of data, Contributed unpublished essential data or reagents; OB, Drafting or revising the article, Contributed unpublished essential data or reagents; MD, XD, Conception and design, Analysis and interpretation of data, Drafting or revising the article The authors declare that no competing interests exist.

Attached Files

Published - elife-02230-v2.pdf

Supplemental Material - elife-02230-media1.mp4

Supplemental Material - elife-02230-media2.mp4

Supplemental Material - elife-02230-media3.mp4

Supplemental Material - elife-02230-media4.mp4

Supplemental Material - elife-02230-media5.mp4

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elife-02230-media1.mp4

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Additional details

Identifiers

PMCID
PMC4095940
Eprint ID
106675
Resolver ID
CaltechAUTHORS:20201113-163228551

Related works

Describes
10.7554/eLife.02230.007 (DOI)
10.7554/eLife.02230.008 (DOI)
10.7554/eLife.02230.009 (DOI)
10.7554/eLife.02230.010 (DOI)
10.7554/eLife.02230.011 (DOI)

Funding

Fondation pour la Recherche Médicale
Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)
Agence Nationale pour la Recherche (ANR)
PCV DYNAFT 08-PCVI-0013
Agence Nationale pour la Recherche (ANR)
DynamIC ANR-12-BSV5-0018
Nikon France

Dates

Created
2020-11-25
Created from EPrint's datestamp field
Updated
2023-06-02
Created from EPrint's last_modified field