BtuB-Dependent Infection of the T5-like Yersinia Phage ϕR2-01.

Lotta J Happonen,Maria I Pajunen,Mikael Skurnik,Jin Woo Jun

doi:10.3390/v13112171

Lotta J Happonen, Maria I Pajunen + Show 2 more

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https://doi.org/10.3390/v13112171

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Abstract

Yersinia enterocolitica is a food-borne Gram-negative pathogen responsible for several gastrointestinal disorders. Host-specific lytic bacteriophages have been increasingly used recently as an alternative or complementary treatment to combat bacterial infections, especially when antibiotics fail. Here, we describe the proteogenomic characterization and host receptor identification of the siphovirus vB_YenS_ϕR2-01 (in short, ϕR2-01) that infects strains of several Yersinia enterocolitica serotypes. The ϕR2-01 genome contains 154 predicted genes, 117 of which encode products that are homologous to those of Escherichia bacteriophage T5. The ϕR2-01 and T5 genomes are largely syntenic, with the major differences residing in areas encoding hypothetical ϕR2-01 proteins. Label-free mass-spectrometry-based proteomics confirmed the expression of 90 of the ϕR2-01 genes, with 88 of these being either phage particle structural or phage-particle-associated proteins. In vitro transposon-based host mutagenesis and ϕR2-01 adsorption experiments identified the outer membrane vitamin B12 receptor BtuB as the host receptor. This study provides a proteogenomic characterization of a T5-type bacteriophage and identifies specific Y. enterocolitica strains sensitive to infection with possible future applications of ϕR2-01 as a food biocontrol or phage therapy agent.

Highlights

Yersinia enterocolitica is a zoonotic, food-borne Gram-negative bacterium of the familyEnterobacteriaceae that can cause yersiniosis in humans and animals [1]
In addition to T5, we identified several other closely related viruses based on similarity searches (Table S4), including among others the siphophage Stitch [56], Salmonella phage Shivani [57], Escherichia phage vB_EcoS_FFH1 [58], Salmonella phage OSY-STA [59], Salmonella phage Seabear [60], Salmonella phage Seafire [61], Salmonella phage STG2 [62], In total, 117 of the 154 predicted φR2-01 genes have unique counterparts in T5 and, in contrast, 32 of the φR2-01 genes are missing in T5 (Table S3)
Lytic bacteriophages are powerful tools to be used in phage therapy and as biocontrol agents

Summary

Introduction

Yersinia enterocolitica is a zoonotic, food-borne Gram-negative bacterium of the familyEnterobacteriaceae that can cause yersiniosis in humans and animals [1]. The main animal reservoir for Y. enterocolitica is pigs, and pork-derived products are thought to be the main source of human infections, in addition to blood transfusions and intake of contaminated drinking water [1,2]. Several bacteriophages that infect Y. enterocolitica have previously been isolated from the raw incoming sewage of city sewage treatment plants in Finland and from pig stool samples [5,6,7]. These phages were isolated using different host strains for enrichment. Many of these bacteriophages use different parts of the Y. enterocolitica lipopolysaccharide (LPS) as the receptor [5,7]

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