sbcB sbcC null mutations allow RecF-mediated repair of arrested replication forks in rep recBC mutants.
We have proposed previously that, in Escherichia coli, blockage of replication forks can lead to the reversal of the fork. Annealing of the newly synthesized strands creates a double-stranded end adjacent to a Holliday junction. The junction is migrated away from the DNA end by RuvAB and can be cleaved by RuvC, while RecBCD is required for the repair of the double-stranded tail. Consequently, the rep mutant, in which replication arrests are frequent and fork reversal occurs, requires RecBCD for growth. We show here that the combination of sbcB sbcCD null mutations restores the viability to rep recBC mutants by activation of the RecF pathway of recombination. This shows that the proteins belonging to the RecF pathway are able to process the DNA ends made by the replication fork reversal into a structure that allows recombination-dependent replication restart. However, we confirm that, unlike sbcB null mutations, sbcB15, which suppresses all other recBC mutant defects, does not restore the viability of rep recBC sbcCD strains. We also show that ruvAB inactivation suppresses the lethality and the formation of double-stranded breaks (DSBs) in a rep recBC recF strain, totally deficient for homologous recombination, as well as in rep recBC mutants. This confirms that RuvAB processing of arrested replication forks is independent of the presence of recombination intermediates.
Bidnenko
V
V
Génétique Microbienne, Institut National de la Recherche Agronomique, 78352 Jouy en Josas Cedex, France.
Seigneur
M
M
Penel-Colin
M
M
Bouton
M F
MF
Dusko Ehrlich
S
S
Michel
B
B
D001426:Bacterial Proteins / Q000235:genetics*; D004249:DNA Damage / Q000235:genetics; D004261:DNA Replication / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D003851:Deoxyribonucleases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000235:genetics* / Q000378:metabolism*; D005798:Genes, Bacterial; D005838:Genotype; D009154:Mutation; D011995:Recombination, Genetic; D013489:Suppression, Genetic; D013696:Temperature
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C061563:SbcC protein, Bacteria; C489229:SbcC protein, E coli; C489340:recF protein, E coli; C066218:recF protein, Bacteria; D003851:Deoxyribonucleases; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V; C468322:exodeoxyribonuclease V, E coli