Decreased Host-Cell ATP Levels Affects Bacteriophage Replication in Knockout E. coli Strains

Abstract

Bacteriophages are viruses that use host cell metabolic resources for replication. Altering Escherichia coli's ATP production pathway can inhibit bacteriophage replication, offering a new approach to bacteriophage therapy.The atp genes encode ATP synthase subunits crucial for ATP generation in E. coli. Knockouts ΔatpA, B, D, E, and H, alongside the parent strain, were studied. Focus narrowed to ΔatpA and B due to significant deviations from the parent strain. It is hypothesized that these knockout strains reduce growth in E. coli and bacteriophage due to decreased ATP production, vital for metabolism and phage replication. Comparative growth assays of E. coli parent and ATP knockout strains were conducted in LB-rich media and M9 minimal media. T4 bacteriophage replication was measured through lysis curves, plaque assays, and two time-point phage titer experiments, chosen for consistent replication. Characterization of T4 bacteriophage replication revealed ΔatpA's crucial role, showing difficulties in growth and lysing. ΔatpA required 10-4 dilutions in phage titer experiments due to low PFU/mL, contrasting with 10-7 dilutions for other strains. ATP assay data showed significantly lower ATP concentration (319nM) in ΔatpB compared to the parent strain, also implying its crucial role in ATP synthesis.Future research will focus on characterizing phage replication in ATP synthase knockout strains using E. coli ATP synthase inhibitors to deepen understanding of phage-host interactions. Controlled bacteriophage manipulation can be studied further to have a better understanding of the application of bacteriophage therapy and to potentially improve its clinical efficacy.