Single molecule microscopy reveals key physical features of repair foci in living cells.
DNA repair
S. cerevisiae
liquid-liquid phase separation
nuclear sub-compartments
physics of living systems
single molecule microscopy
single particle tracking
Journal
eLife
ISSN: 2050-084X
Titre abrégé: Elife
Pays: England
ID NLM: 101579614
Informations de publication
Date de publication:
05 02 2021
05 02 2021
Historique:
received:
30
06
2020
accepted:
26
01
2021
pubmed:
6
2
2021
medline:
4
2
2022
entrez:
5
2
2021
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 33543712
doi: 10.7554/eLife.60577
pii: 60577
pmc: PMC7924958
doi:
pii:
Substances chimiques
RAD52 protein, S cerevisiae
0
RFA1 protein, S cerevisiae
0
Rad52 DNA Repair and Recombination Protein
0
Replication Protein A
0
Saccharomyces cerevisiae Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
© 2021, Miné-Hattab et al.
Déclaration de conflit d'intérêts
JM, MH, MV, CG, TM, MD, AT No competing interests declared, AW Reviewing editor, eLife
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