Disruption of the F plasmid partition complex in vivo by partition protein SopA.
The SopA protein plays an essential, though so far undefined, role in partition of the mini-F plasmid but, when overproduced, it causes loss of mini-F from growing cells. Our investigation of this phenomenon has revealed that excess SopA protein reduces the linking number of mini-F. It appears to do so by disturbing the partition complex, in which SopB normally introduces local positive supercoiling upon binding to the sopC centromere, as it occurs only in plasmids carrying sopC and in the presence of SopB protein. SopA-induced reduction in linking number is not associated with altered sop promoter activity or levels of SopB protein and occurs in the absence of changes in overall supercoil density. SopA protein mutated in the ATPase nucleotide-binding site (K120Q) or lacking the presumed SopB interaction domain does not induce the reduction in linking number, suggesting that excess SopA disrupts the partition complex by interacting with SopB to remove positive supercoils in an ATP-dependent manner. Destabilization of mini-F also depends on sopC and SopB, but the K120Q mutant retains some capacity for destabilizing mini-F. SopA-induced destabilization thus appears to be complex and may involve more than one SopA activity. The results are interpreted in terms of a regulatory role for SopA in the oligomerization of SopB dimers bound to the centromere.
Lemonnier
M
M
Laboratoire de Microbiologie et Génétique Moléculaire, 118 Route de Narbonne, 31062 Toulouse, France.
Bouet
J Y
JY
Libante
V
V
Lane
D
D
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D029968:Escherichia coli Proteins; D005144:F Factor / Q000235:genetics* / Q000378:metabolism*; D010641:Phenotype
D001426:Bacterial Proteins; D003470:Culture Media; D029968:Escherichia coli Proteins; C060972:SopB protein, Bacteria; C489223:SopB protein, E coli; C060971:SopA protein, Bacteria