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Abstract

The segregation of the DNA, also called partition for procaryotes, is the process allowing any organisms to transmit its genetic heritage to next generation. In bacteria, mitotic stability of plasmids and many chromosomes depends on replicon-specific systems which comprise a centromere, a centromere-binding protein and an ATPase. We have taken as a model, the lowcopy number plasmid F of Escherichia coli. Centromere-binding protein SopB binds to sopC centromere and forms the partition complex. This nucleoproteic complex is recognized by the SopA “Walker-box” ATPase. SopA shares with other partition ATPase the capacity of self assembly in presence of ATP. This dynamic self-assembly would allow active partition during bacterial division. Previous work in our team showed SopA is also able to bind to non specific DNA in an ATP-dependant manner whereby polymerization is inhibited. Indeed, DNA inhibited this polymerization and cause breakdown of pre-formed polymers. SopB counteracted this DNA effect by binding itself to and masking DNA. We had proposed a model in which the polymerization is spacially regulated. Nucleoid DNA prevent inappropriate SopA polymerization but when SopB is present in high concentration, it create a DNA-depleted zone within SopA can initiate polymerization. The regulation of the dynamic behaviour of the "driving" protein of the system would be necessary for the process of partition. To support our model, we looked for a DNA binding domain in SopA. We have found a SopA mutant, defective for ATP dependent DNA binding. Only the activities of SopA dependent on this binding were affected: the inhibition of the polymerisation is abolished, as the stimulation of the ATPase activity and the intracellular localization. Moreover, this mutant is defective for plasmid stabilization. This last result confirms the implication of the nucleoïd DNA in regulation of the dynamic behavior of SopA, which is necessary for the partition of the plasmide F.