Spatial and temporal organization of RecA in the
DNA damage
DNA repair intermediates
E. coli
RecA
SOS response
live cell imaging
molecular biophysics
physics of living systems
single molecule imaging
structural biology
Journal
eLife
ISSN: 2050-084X
Titre abrégé: Elife
Pays: England
ID NLM: 101579614
Informations de publication
Date de publication:
05 02 2019
05 02 2019
Historique:
received:
18
10
2018
accepted:
22
01
2019
entrez:
6
2
2019
pubmed:
6
2
2019
medline:
27
3
2020
Statut:
epublish
Résumé
The RecA protein orchestrates the cellular response to DNA damage via its multiple roles in the bacterial SOS response. Lack of tools that provide unambiguous access to the various RecA states within the cell have prevented understanding of the spatial and temporal changes in RecA structure/function that underlie control of the damage response. Here, we develop a monomeric C-terminal fragment of the λ repressor as a novel fluorescent probe that specifically interacts with RecA filaments on single-stranded DNA (RecA*). Single-molecule imaging techniques in live cells demonstrate that RecA is largely sequestered in storage structures during normal metabolism. Upon DNA damage, the storage structures dissolve and the cytosolic pool of RecA rapidly nucleates to form early SOS-signaling complexes, maturing into DNA-bound RecA bundles at later time points. Both before and after SOS induction, RecA* largely appears at locations distal from replisomes. Upon completion of repair, RecA storage structures reform.
Identifiants
pubmed: 30717823
doi: 10.7554/eLife.42761
pii: 42761
pmc: PMC6363387
doi:
pii:
Substances chimiques
DNA, Bacterial
0
DNA-Binding Proteins
0
Escherichia coli Proteins
0
recA protein, E coli
0
Rec A Recombinases
EC 2.7.7.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIGMS NIH HHS
ID : U01 GM032335
Pays : United States
Organisme : NIH HHS
ID : GM32335
Pays : United States
Organisme : NIH HHS
ID : U01 GM32335
Pays : United States
Organisme : Australian Research Council
ID : DP150100956
Pays : International
Organisme : Australian Research Council
ID : DP180100858
Pays : International
Organisme : Australian Research Council
ID : FL140100027
Pays : International
Organisme : NIH HHS
ID : Intramural Research Program
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM032335
Pays : United States
Déclaration de conflit d'intérêts
HG, BP, JL, SJ, KG, ZR, EW, RW, MC, Av No competing interests declared
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