TPM analyses reveal that FtsK contributes both to the assembly and the activation of the XerCD-dif recombination synapse.
Circular chromosomes can form dimers during replication and failure to resolve those into monomers prevents chromosome segregation, which leads to cell death. Dimer resolution is catalysed by a highly conserved site-specific recombination system, called XerCD-dif in Escherichia coli. Recombination is activated by the DNA translocase FtsK, which is associated with the division septum, and is thought to contribute to the assembly of the XerCD-dif synapse. In our study, direct observation of the assembly of the XerCD-dif synapse, which had previously eluded other methods, was made possible by the use of Tethered Particle Motion, a single molecule approach. We show that XerC, XerD and two dif sites suffice for the assembly of XerCD-dif synapses in absence of FtsK, but lead to inactive XerCD-dif synapses. We also show that the presence of the γ domain of FtsK increases the rate of synapse formation and convert them into active synapses where recombination occurs. Our results represent the first direct observation of the formation of the XerCD-dif recombination synapse and its activation by FtsK.
CNRS; IPBS (Institut de Pharmacologie et de Biologie Structurale); 205 route de Narbonne BP 64182, F-31077 Toulouse, France, Université de Toulouse; UPS; IPBS; F-31077 Toulouse, France, Université de Toulouse; UPS; LMGM (Laboratoire de Microbiologie et Génétique Moléculaires); F-31062 Toulouse, France and CNRS; LMGM; F-31062 Toulouse, France.
Cheikh Tidiane
CT
Diagne
Maya
M
Salhi
Estelle
E
Crozat
Laurence
L
Salomé
Francois
F
Cornet
Philippe
P
Rousseau
Catherine
C
Tardin
D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D019426:Integrases / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D009038:Motion; D017434:Protein Structure, Tertiary; D011995:Recombination, Genetic
D029968:Escherichia coli Proteins; C096452:FtsK protein, E coli; D008565:Membrane Proteins; C473360:XerC protein, E coli; D019426:Integrases; C473361:XerD protein, E coli