ParBF DNA binding profiles from variants of the CTP binding motif

We report the genome-wide analysis from chromatin immunoprecipitated DNA (ChIP-sequencing) of the DNA binding pattern of ParBF (SopB) of plasmid F. This study, performed in E. coli, investigates the impact of mutations in the CTP binding motif BoxIII on ParBF DNA binding profiles in vivo. We found that ParBF-N153A allele binds to parS as WT but is impaired in clamp formation thus abolishing the long distance DNA binding on parS-proximal DNA. By contrast, ParBF-R156A DNA binding profile reveals that it binds to parS, converts to clamps that diffuse over several Kb and unbinds non-specific DNA in a similar way as WT. However, the reduction in ParB binding at proximity of parS revealed that ParBF-R156A is impaired in ParB-ParB interaction, thus preventing the assembly of ParB condensates. Overall design: Chromatin immunoprecipitation DNA-sequencing for ParB of plasmid F (ParBF) in Escherichia coli. Biological replicates are labelled IP1 and IP2. Anti ParBF antibody is affinity purified before used.