Genome-wide overexpression screen of Pseudomonas putida phage Emajogi

Abstract

Bacteriophage genomes contain large fractions of uncharacterized genes, motivating high-throughput functional screens to illuminate their roles in host interaction and cytotoxicity. Here, we performed a genome wide overexpression assay for a Pseudomonas putida KT2440 infecting phage, Emajogi, and identified eight cytotoxic genes that reduce host growth under both basal and induced expression. We show that these proteins are conserved across phages that infect a diverse set of hosts. The most cytotoxic gene identified was a hypothetical protein (gp18) with no predicted functional domains. Transcriptomic profiling revealed that overexpression of gp18 caused a pronounced metabolic remodeling of the cell, likely shifting cell resources to phage production. Overall, this genome wide screen provides information about phage genes that were previously unannotated that can support biotechnology and novel antimicrobial development.