The study provides valuable insights into the role of DciA in bacterial replication through LLPS-mediated condensates. However, certain aspects would benefit from further clarification:
– Could the authors discuss how the ionic conditions used in the experiments (e.g., high NaCl concentrations) compare to the native bacterial intracellular environment? How might this influence DciA’s phase-separation behavior in vivo?
– The FRAP and FIDA analyses effectively characterize droplet dynamics, but could additional experiments, such as cross-linking or structural studies, help identify the specific residues or domains of DciA involved in driving LLPS?
– Given the observed differences between DciA- and DnaC-mediated LLPS, do the authors hypothesize that these mechanisms confer distinct functional advantages in replication initiation across bacterial species?