Movement and equipositioning of plasmids by ParA filament disassembly.
Bacterial plasmids encode partitioning (par) loci that confer stable plasmid inheritance. We showed previously that, in the presence of ParB and parC encoded by the par2 locus of plasmid pB171, ParA formed cytoskeletal-like structures that dynamically relocated over the nucleoid. Simultaneously, the par2 locus distributed plasmids regularly over the nucleoid. We show here that the dynamic ParA patterns are not simple oscillations. Rather, ParA nucleates and polymerizes in between plasmids. When a ParA assembly reaches a plasmid, the assembly reaction reverses into disassembly. Strikingly, plasmids consistently migrate behind disassembling ParA cytoskeletal structures, suggesting that ParA filaments pull plasmids by depolymerization. The perpetual cycles of ParA assembly and disassembly result in continuous relocation of plasmids, which, on time averaging, results in equidistribution of the plasmids. Mathematical modeling of ParA and plasmid dynamics support these interpretations. Mutational analysis supports a molecular mechanism in which the ParB/parC complex controls ParA filament depolymerization.
Ringgaard
Simon
S
Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne NE2 4HH, United Kingdom.
van Zon
Jeroen
J
Howard
Martin
M
Gerdes
Kenn
K
D019943:Amino Acid Substitution; D001419:Bacteria / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011108:Polymers / Q000378:metabolism; D016963:Thermus thermophilus / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D011108:Polymers; C052883:chromosome partition proteins, bacterial; D049452:Green Fluorescent Proteins