1887

Abstract

ParA belongs to a large subfamily of Walker-type ATPases acting as partitioning proteins in bacteria. ParA has the ability to both self-associate and interact with its partner ParB. Analysis of the deletion mutants defined the part of the protein involved in dimerization and interactions with ParB. Here, a set of ParA alanine substitution mutants in the region between E67 and L85 was created and analysed and . All mutants impaired in dimerization (substitutions at positions M74, H79, Y82 and L84) were also defective in interactions with ParB, suggesting that ParA–ParB interactions depend on the ability of ParA to dimerize. Mutants with alanine substitutions at positions E67, C68, L70, E72, F76, Q83 and L85 were not impaired in dimerization, but were defective in interactions with ParB. The dimerization interface partly overlapped the pseudo-hairpin, involved in interactions with ParB. ParA mutant derivatives tested showed no defects in ATPase activity. Two alleles (, whose product can neither self-interact nor interact with ParB, and , whose product is impaired in interactions with ParB, but not in dimerization) were introduced into the chromosome by homologous gene exchange. Both mutants showed defective separation of ParB foci, but to different extents. Only PAO1161 was visibly impaired in terms of chromosome segregation, growth rate and motility, similar to a -null mutant.

Funding
This study was supported by the:
  • Wellcome Trust Collaborative Research Initiative (Award 067068/Z/02/Z and 056022/Z/98/Z)
  • MNiSW (Award 2913/B/PO1/2008/34)
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2014-11-01
2021-10-25
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