Comparative Genomics and Characterization of the Late Promoter pR’ from Shiga Toxin Prophages in Escherichia coli

Shiga-toxin producing Escherichia coli (STEC) causes human illness ranging from mild diarrhea to death. The bacteriophage encoded stx genes are located in the late transcription region, downstream of the antiterminator Q. The transcription of the stx genes is directly under the control of the late p...

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Bibliographic Details
Published in:Viruses 2018-10, Vol.10 (11), p.595
Main Authors: Zhang, Ling, Simpson, David, McMullen, Lynn, Gänzle, Michael
Format: Article
Language:English
Subjects:
Diarrhea
E coli
Epidemics
Escherichia coli
Flow cytometry
Genes
Genomes
genomic characterization
Genomics
microscope
Mitomycin C
Nucleotide sequence
phage induction efficiency
Phages
Phylogenetics
Phylogeny
Plasmids
Prophages
Proteins
Reporter gene
RNA polymerase
sequence diversity
Shiga toxin
Shiga toxin prophage
Stx gene
Transcription
Virulence
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Summary:Shiga-toxin producing Escherichia coli (STEC) causes human illness ranging from mild diarrhea to death. The bacteriophage encoded stx genes are located in the late transcription region, downstream of the antiterminator Q. The transcription of the stx genes is directly under the control of the late promoter pR’, thus the sequence diversity of the region between Q and stx, here termed the pR’ region, may affect Stx toxin production. Here, we compared the gene structure of the pR’ region and the stx subtypes of nineteen STECs. The sequence alignment and phylogenetic analysis suggested that the pR’ region tends to be more heterogeneous than the promoter itself, even if the prophages harbor the same stx subtype. Furthermore, we established and validated transcriptional fusions of the pR’ region to the DsRed reporter gene using mitomycin C (MMC) induction. Finally, these constructs were transformed into native and non-native strains and examined with flow cytometry. The results showed that induction levels changed when pR’ regions were placed under different regulatory systems. Moreover, not every stx gene could be induced in its native host bacteria. In addition to the functional genes, the diversity of the pR’ region plays an important role in determining the level of toxin induction.
ISSN:1999-4915
1999-4915
DOI:10.3390/v10110595