Abstract

Salmonella enterica serovar Newport bacteriophage 7-11 shares 41 homologous ORFs with Escherichia coli phage phiEco32, and both phages encode a protein similar to bacterial RNA polymerase promoter specificity σ subunit. Here, we investigated the temporal pattern of 7-11 gene expression during infection and compared it to the previously determined transcription strategy of phiEco32. Using primer extension and in vitro transcription assays, we identified eight promoters recognized by host RNA polymerase holoenzyme containing 7-11 σ subunit SaPh711_gp47. These promoters are characterized by a bipartite consensus, GTAAtg-(16)-aCTA, and are located upstream of late phage genes. While dissimilar from single-element middle and late promoters of phiEco32 recognized by holoenzymes formed by the phi32_gp36 σ factor, the 7-11 late promoters are located at genome positions similar to those of phiEco32 middle and late promoters. Two early 7-11 promoters are recognized by the RNA polymerase holoenzyme containing the host primary σ70 factor. Unlike the case of phiEco32, no shut-off of σ70-dependent transcription is observed during 7-11 infection and there are no middle promoters. These differences can be explained by the fact that phage 7-11 does not encode a homologue of phi32_gp79, an inhibitor of host and early phage transcription and an activator of transcription by the phi32_gp36-holoenzyme.

Highlights

  • Salmonella enterica serovar Newport phage 7-11 is a podovirus with a distinctive, strongly elongated virion head [1]

  • PhiEco32 remains the best-studied member of the Kuravirus genus to date. It relies on host RNA polymerase (RNAP) for its development within the host cell [6–8]

  • Over one hundred bacteriophages with similarities in genomic sequence and gene organization were submitted to public databases, including, among others, E. coli (APEC) phages NJ01 (NC_018835), ECB2 (NC_018859), SU10 (KM044272) and 172-1 (KP308307) [3–5]

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Summary

Introduction

Salmonella enterica serovar Newport phage 7-11 is a podovirus with a distinctive, strongly elongated virion head [1]. PhiEco remains the best-studied member of the Kuravirus genus to date. As most bacteriophages, it relies on host RNA polymerase (RNAP) for its development within the host cell [6–8]. Phage genes are transcribed from four promoters located at the beginning of the early gene cluster. These promoters are recognized by the housekeeping σ70 RNAP holoenzyme of the host. All late genes and at least some of the middle genes of phiEco are transcribed from promoters recognized by an RNAP holoenzyme containing phage σ factor phi32_gp. A small phage polypeptide, phi32_gp, a product of a middle gene, inhibits transcription by the σ70 holoenzyme and stimulates phi32_gp36-dependent transcription in vitro [2,6]. Phi32_gp may be responsible for regulated expression of phage genes and host transcription shut-off

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