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From Single-Molecule Interactions to Population-Level Dynamics: Understanding the Complex Organization of RNA Pol II in the Nucleus of Living Cells

Cissé, Ibrahim I. and Izeddin, Ignacio and Causse, Sebastien and El Beheiry, Mohamed and Dugast-Darzacq, Claire and Muresan, Leila and Récamier, Vincent and Dahan, Maxime and Darzacq, Xavier (2012) From Single-Molecule Interactions to Population-Level Dynamics: Understanding the Complex Organization of RNA Pol II in the Nucleus of Living Cells. Biophysical Journal, 102 (3, S1). 475a. ISSN 0006-3495. doi:10.1016/j.bpj.2011.11.2607. https://resolver.caltech.edu/CaltechAUTHORS:20201211-154706035

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Abstract

Transcription involves a complex exchange within a reservoir of proteins in the nucleoplasm, and the specific recruitment of individual proteins at specific gene loci. However, understanding the spatial distribution of individual proteins and the temporal behavior in the nucleus of living cells remains challenging. Using 3D super-resolution fluorescence microscopy and cluster analysis, we observe that the distribution of RNA Polymerase II (Pol II) cluster sizes, measured as the number of polymerases per cluster, follows a −3/2 power law. Radial dependent analysis of the spatial distribution of Pol II also shows scale-invariance, consistent with a so-called self-organized criticality in a fractal geometry of dimension ∼2.7. These results suggest a diffusion-based mechanism whereby, via transient interactions, massive recruitment and dismissal of pol II molecules can occur at specific loci in the nucleoplasm. Kinetic measurements using single-molecule detection in live cells reveal Pol II binding dynamics within minutes. Serum-induced transcription increased Pol II binding kinetics in live cells by an order of magnitude. Together, these results provide a comprehensive view of the spatio-temporal organization of Pol II in the nucleus: from the global population distribution, to single molecule recruitment at specific loci in live cells. This comprehensive single-cell approach can be adopted for other proteins beside RNA Pol II, for real-time quantification of protein organization in vivo, with single-molecule sensitivity.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.bpj.2011.11.2607DOIArticle
ORCID:
AuthorORCID
Cissé, Ibrahim I.0000-0002-8764-1809
Izeddin, Ignacio0000-0002-8476-3915
Causse, Sebastien0000-0003-3246-3491
El Beheiry, Mohamed0000-0001-9827-6619
Dugast-Darzacq, Claire0000-0001-8602-3534
Muresan, Leila0000-0002-7602-0249
Dahan, Maxime0000-0002-4943-2491
Darzacq, Xavier0000-0003-2537-8395
Additional Information:© 2011 Biophysical Society. Published by Elsevier Inc. Available online 14 February 2012.
Issue or Number:3, S1
DOI:10.1016/j.bpj.2011.11.2607
Record Number:CaltechAUTHORS:20201211-154706035
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20201211-154706035
Official Citation:Ibrahim I. Cisse, Ignacio Izeddin, Sebastien Causse, Mohamed El Beheiry, Claire Dugast-Darzacq, Leila Muresan, Vincent Recamier, Maxime Dahan, Xavier Darzacq, From Single-Molecule Interactions to Population-Level Dynamics: Understanding the Complex Organization of RNA Pol II in the Nucleus of Living Cells, Biophysical Journal, Volume 102, Issue 3, Supplement 1, 2012, Page 475a, ISSN 0006-3495, https://doi.org/10.1016/j.bpj.2011.11.2607. (http://www.sciencedirect.com/science/article/pii/S0006349511039555)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:107044
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:11 Dec 2020 23:56
Last Modified:16 Nov 2021 18:59

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