The prophages of Citrobacter rodentium represent a conserved family of horizontally-acquired mobile genetic elements associated with enteric evolution towards pathogenicity
Prophage mediated horizontal gene transfer (HGT) plays a key role in the evolution of bacteria, enabling access to new environmental niches, including pathogenicity. Citrobacter rodentium is a host-adapted intestinal mouse pathogen and important model organism for attaching and effacing (A/E) pathogens including the clinically significant enterohaemorrhagic (EHEC) and enteropathogenic (EPEC) Escherichia coli. Despite containing ten prophage genomic regions, including an active temperate phage, ΦNP, little was known regarding the nature of C. rodentium prophages in the bacterium’s evolution towards pathogenicity. In this study, our characterization of ΦNP led to the discovery of a second, fully functional temperate phage, named ΦSM. We identify the bacterial host-receptor for both phages as lipopolysaccharide (LPS). ΦNP and ΦSM are likely important mediators of HGT in C. rodentium. Bioinformatic analysis of the ten prophage regions reveals cargo genes encoding known virulence factors, including several Type III secretion system (T3SS) effectors. C. rodentium prophages are conserved across a wide range of pathogenic enteric bacteria, including EPEC and EHEC as well as pathogenic strains of Salmonella enterica, Shigella boydii, and Klebsiella pneumoniae. Phylogenetic analysis of core enteric backbone genes compared against prophage evolutionary models suggests that these prophages represent an important, conserved family of horizontally acquired enteric-associated pathogenicity determinants. In addition to highlighting the transformative role of bacteriophage mediated HGT in C. rodentium’s evolution towards pathogenicity, these data suggest that the examination of conserved families of prophages in other pathogenic bacteria and disease outbreaks might provide deeper evolutionary and pathological insights otherwise obscured by more classical analysis.
Biotechnology and Biological Sciences Research Council (BB/H002677/1)