Transcriptomic Analysis of the Campylobacter jejuni Response to T4-Like Phage NCTC 12673 Infection.

Jessica Sacher,Christine Szymanski,James Butcher,Hayley Reynolds,Bob Blasdel,Annika Flint,Rob Lavigne,Alain Stintzi

doi:10.3390/v10060332

Jessica Sacher, Christine Szymanski + Show 6 more

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https://doi.org/10.3390/v10060332

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Abstract

Campylobacter jejuni is a frequent foodborne pathogen of humans. As C. jejuni infections commonly arise from contaminated poultry, phage treatments have been proposed to reduce the C. jejuni load on farms to prevent human infections. While a prior report documented the transcriptome of C. jejuni phages during the carrier state life cycle, transcriptomic analysis of a lytic C. jejuni phage infection has not been reported. We used RNA-sequencing to profile the infection of C. jejuni NCTC 11168 by the lytic T4-like myovirus NCTC 12673. Interestingly, we found that the most highly upregulated host genes upon infection make up an uncharacterized operon (cj0423–cj0425), which includes genes with similarity to T4 superinfection exclusion and antitoxin genes. Other significantly upregulated genes include those involved in oxidative stress defense and the Campylobacter multidrug efflux pump (CmeABC). We found that phage infectivity is altered by mutagenesis of the oxidative stress defense genes catalase (katA), alkyl-hydroxyperoxidase (ahpC), and superoxide dismutase (sodB), and by mutagenesis of the efflux pump genes cmeA and cmeB. This suggests a role for these gene products in phage infection. Together, our results shed light on the phage-host dynamics of an important foodborne pathogen during lytic infection by a T4-like phage.

Highlights

Bacteriophages, the viruses that infect bacteria, represent a diverse class of natural bacterial predators that have shaped bacterial evolution for an estimated three billion years
We found that the host promoters and Rho-independent terminators predicted by [25] could be motifs using Find Individual Motif Occurrences (FIMO), and additional examples were annotated validated by RNA-Seq data, though most transcription appears to be driven by one of two novel A/T
National Collection of Type Cultures (NCTC) 12673 Phage Infection Resulted in Clear Differences in Overall C. jejuni Gene Transcription

Summary

Introduction

Bacteriophages (phages), the viruses that infect bacteria, represent a diverse class of natural bacterial predators that have shaped bacterial evolution for an estimated three billion years. As antibiotic resistant infections threaten to cause more deaths per year than cancer by 2050 [1], there is a great need to explore alternative antimicrobial therapies. Phages represent a viable antibiotic alternative, both in human healthcare settings and in agriculture [2,3,4]. To ensure the safety and efficacy of employing phages in these settings, only well-characterized phages should be used. Since the discovery of phages more than a century ago, phage-host characterization has progressed through many stages, from examining plaque morphology, to molecular genetics using model

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