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Among the mechanisms that control chromosome segregation in bacteria are highly-conserved partitioning systems comprising three components: ParA protein (a deviant Walker-type ATPase), ParB protein (a DNA-binding element) and multiple cis-acting palindromic centromere-like sequences, designated  parS . Ten putative  parS  sites have been identified in the  P .  aeruginosa  PAO1 genome, four localized in close proximity of  oriC  and six, diverged by more than one nucleotide from a perfect palindromic sequence, dispersed along the chromosome. Here, we constructed and analyzed  P .  aeruginosa  mutants deprived of each single  parS  sequence and their different combinations. The analysis included evaluation of a set of phenotypic features, chromosome segregation, and ParB localization in the cells. It was found that ParB binds specifically to all ten  parS  sites, although with different affinities. The  P .  aeruginosa parS  mutant with all ten  parS  sites modified ( parS   null ) is viable however it demonstrates the phenotype characteristic for  parA   null  or  parB   null  mutants: slightly slower growth rate, high frequency of anucleate cells, and defects in motility. The genomic position and sequence of  parS  determine its role in  P .  aeruginosa  biology. It transpired that any one of the four  parS  sites proximal to  oriC  ( parS1  to  parS4 ), which are bound by ParB with the highest affinity, is necessary and sufficient for the  parABS  role in chromosome partitioning. When all these four sites are mutated simultaneously, the strain shows the  parS   null  phenotype, which indicates that none of the remaining six  parS  sites can substitute for these four  oriC -proximal sites in this function. A single ectopic  parS2  (inserted opposite  oriC  in the  parS   null  mutant) facilitates ParB organization into regularly spaced condensed foci and reverses some of the mutant phenotypes but is not sufficient for accurate chromosome segregation.

plos one

A Single parS Sequence from the Cluster of Four Sites Closest to oriC Is Necessary and Sufficient for Proper Chromosome Segregation in Pseudomonas aeruginosa