Single molecule microscopy reveals key physical features of repair foci in living cells
- Institut Curie, France
- Ecole Normale Supérieure, France
- École Normale Supérieure, France
Abstract
In response to double strand breaks (DSB), repair proteins accumulate at damaged sites, forming membrane-less sub-compartments or foci. Here we explored the physical nature of these foci, using single molecule microscopy in living cells. Rad52, the functional homolog of BRCA2 in yeast, accumulates at DSB sites and diffuses ~6 times faster within repair foci than the focus itself, exhibiting confined motion. The Rad52 confinement radius coincides with the focus size: foci resulting from 2 DSBs are twice larger in volume that the ones induced by a unique DSB and the Rad52 confinement radius scales accordingly. In contrast, molecules of the single strand binding protein Rfa1 follow anomalous diffusion similar to the focus itself or damaged chromatin. We conclude that while most Rfa1 molecules are bound to the ssDNA, Rad52 molecules are free to explore the entire focus reflecting the existence of a liquid droplet around damaged DNA.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. Source data files are available on zenodo using the following link: https://zenodo.org/record/4495116.
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Single molecule microscopy reveals key physical features of repair foci in living cellsZenodo, doi: 10.5281/zenodo.4495116.
Article and author information
Author details
Judith Miné-Hattab
Aleksandra M Walczak
Funding
Agence Nationale de la Recherche (ANR-11-LABEX-0044 DEEP)
- Angela Taddei
Agence Nationale de la Recherche (ANR-10-IDEX-0001-02 PSL)
- Angela Taddei
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2021, Miné-Hattab et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Single molecule microscopy reveals key physical features of repair foci in living cells
eLife 10:e60577.
https://doi.org/10.7554/eLife.60577
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