Effect of a Defective Clamp Loader Complex of DNA Polymerase III on Growth and SOS Response in Pseudomonas aeruginosa

DNA polymerase III (Pol III) is the replicative enzyme in bacteria. It consists of three subcomplexes, the catalytic core, the beta clamp, and the clamp loader. While this complex has been thoroughly characterized in the model organism Escherichia coli, much less is known about its functioning and/or its specific properties in other bacteria. Biochemical studies highlighted specific features in the clamp loader subunit psi of Pseudomonas aeruginosa as compared to its E. coli counterpart, and transposon mutagenesis projects identified the psi-encoding gene holD among the strictly essential core genes of P. aeruginosa. By generating a P. aeruginosa holD conditional mutant, here we demonstrate that, as previously observed for E. coli holD mutants, HolD-depleted P. aeruginosa cells show strongly decreased growth, induction of the SOS response, and emergence of suppressor mutants at high frequency. However, differently from what was observed in E. coli, the growth of P. aeruginosa cells lacking HolD cannot be rescued by the deletion of genes for specialized DNA polymerases. We also observed that the residual growth of HolD-depleted cells is strictly dependent on homologous recombination functions, suggesting that recombination-mediated rescue of stalled replication forks is crucial to support replication by a psi-deficient Pol III enzyme in P. aeruginosa.

Automated summary

DNA polymerase III is an important replicative enzyme in bacteria. This study investigates the specific properties and functioning of DNA polymerase III in Pseudomonas aeruginosa. The findings show that a conditional mutant lacking the HolD subunit of DNA polymerase III in P. aeruginosa exhibits decreased growth, induction of the SOS response, and emergence of suppressor mutants. Interestingly, unlike in Escherichia coli, the growth of P. aeruginosa cells lacking HolD cannot be rescued by deleting specialized DNA polymerase genes. The study also highlights the essential role of homologous recombination in supporting replication by a psi-deficient Pol III enzyme in P. aeruginosa.