Real-Time Dynamics of RNA Polymerase II Clustering in Live Human Cells
Abstract
Transcription is reported to be spatially compartmentalized in nuclear transcription factories with clusters of RNA polymerase II (Pol II). However, little is known about when these foci assemble or their relative stability. We developed a quantitative single-cell approach to characterize protein spatiotemporal organization, with single-molecule sensitivity in live eukaryotic cells. We observed that Pol II clusters form transiently, with an average lifetime of 5.1 (± 0.4) seconds, which refutes the notion that they are statically assembled substructures. Stimuli affecting transcription yielded orders-of-magnitude changes in the dynamics of Pol II clusters, which implies that clustering is regulated and plays a role in the cell's ability to effect rapid response to external signals. Our results suggest that transient crowding of enzymes may aid in rate-limiting steps of gene regulation.
Additional Information
© 2013, American Association for the Advancement of Science. Received for publication April 11, 2013. Accepted for publication June 17, 2013. We acknowledge J. T. Lis, I. Golding, R. Phillips, O. Bensaude, and E. Bertrand for valuable discussions. We thank members of the Darzacq, Dahan, and Howard Hughes Medical Institute–Janelia Farm Transcription Imaging Consortium groups for suggestions. I.I.C. was supported by fellowships from the European Molecular Biology Organization and the Foundation Pierre Gilles de Gennes (FPGG). I.I. acknowledges the Netherlands Organization for Scientific Research and FPGG for financial support. A.S. was supported by La Ligue nationale contre le cancer. L.M. acknowledges support from Centre de Génétique Moléculaire UPR 3404. This work was supported by grants Agence Nationale de la Recherche Pol2Kinetics to X.D. and DYNAFT to X.D. and M.D.; X.D. and M.D. acknowledge the support of Nikon France.Errata
Revised 9 August 2013: The full references are listed, and the numbering for the supplementary references has been corrected. The supplementary text and the methods section have been correctly rearranged.Attached Files
Supplemental Material - 1239053.Cisse.SM.revision1.pdf
Files
| Name | Size | Download all |
|---|---|---|
|
md5:5ca979156be3c63197108ce73dea021a
|
1.3 MB | Preview Download |
Additional details
- Eprint ID
- 106658
- Resolver ID
- CaltechAUTHORS:20201112-143949341
- European Molecular Biology Organization (EMBO)
- Fondation Pierre Gilles de Gennes
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)
- Ligue Nationale Contre le Cancer
- UPR 3404
- Centre de Génétique Moléculaire
- Pol2Kinetics
- Agence Nationale pour la Recherche (ANR)
- DYNAFT
- Agence Nationale pour la Recherche (ANR)
- Nikon France
- Created
-
2020-11-16Created from EPrint's datestamp field
- Updated
-
2021-11-16Created from EPrint's last_modified field