Concerted action of plasmid maintenance functions: partition complexes create a requirement for dimer resolution.
Partition of prokaryotic DNA requires formation of specific protein-centromere complexes, but an excess of the protein can disrupt segregation. The mechanisms underlying this destabilization are unknown. We have found that destabilization by the F plasmid partition protein, SopB, of plasmids carrying the F centromere, sopC, results from the capacity of the SopB-sopC partition complex to stimulate plasmid multimerization. Mutant SopBs unable to destabilize failed to increase multimerization. Stability of wild-type mini-F, whose ResD/rfsF site-specific recombination system enables it to resolve multimers to monomers, was barely affected by excess SopB. Destabilization of plasmids lacking the rfsF site was suppressed by recF, recO and recR, but not by recB, mutant alleles, indicating that multimerization is initiated from single-strand gaps. SopB did not alter the amounts or distribution of replication intermediates, implying that SopB-DNA complexes do not create single-strand gaps by blocking replication forks. Rather, the results are consistent with SopB-DNA complexes channelling gapped molecules into the RecFOR recombination pathway. We suggest that extended SopB-DNA complexes increase the likelihood of recombination between sibling plasmids by keeping them in close contact prior to SopA-mediated segregation. These results cast plasmid site-specific resolution in a new role - compensation for untoward consequences of partition complex formation.
Bouet
Jean-Yves
JY
Laboratoire de Microbiologie et Génétique Moléculaire, Centre National de Recherche, Scientifique, Faculté Université Paul Sabatier, 118 Route de Narbonne, 31062 Toulouse, France.
Bouvier
Marie
M
Lane
David
D
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020090:Chromosome Segregation; D019281:Dimerization; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000502:physiology*; D011487:Protein Conformation
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C060972:SopB protein, Bacteria; C489223:SopB protein, E coli; C060971:SopA protein, Bacteria