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      Draft Genome Sequences of Five Shiga Toxin-Producing Escherichia coli Isolates Harboring the New and Recently Described Subtilase Cytotoxin Allelic Variant subAB 2-3

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          ABSTRACT

          We present here the draft genome sequences of five Shiga toxin-producing Escherichia coli (STEC) strains which tested positive in a primary subAB screening. Assembly and annotation of the draft genomes revealed that all strains harbored the recently described allelic variant subAB 2-3 . Based on the sequence data, primers were designed to identify and differentiate this variant.

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          A New Family of Potent AB5 Cytotoxins Produced by Shiga Toxigenic Escherichia coli

          Adrienne W Paton, Potjanee Srimanote, Ursula Talbot … (2004)
          The Shiga toxigenic Escherichia coli (STEC) O113:H21 strain 98NK2, which was responsible for an outbreak of hemolytic uremic syndrome, secretes a highly potent and lethal subtilase cytotoxin that is unrelated to any bacterial toxin described to date. It is the prototype of a new family of AB5 toxins, comprising a single 35-kilodalton (kD) A subunit and a pentamer of 13-kD B subunits. The A subunit is a subtilase-like serine protease distantly related to the BA_2875 gene product of Bacillus anthracis. The B subunit is related to a putative exported protein from Yersinia pestis, and binds to a mimic of the ganglioside GM2. Subtilase cytotoxin is encoded by two closely linked, cotranscribed genes (subA and subB), which, in strain 98NK2, are located on a large, conjugative virulence plasmid. Homologues of the genes are present in 32 out of 68 other STEC strains tested. Intraperitoneal injection of purified subtilase cytotoxin was fatal for mice and resulted in extensive microvascular thrombosis, as well as necrosis in the brain, kidneys, and liver. Oral challenge of mice with E. coli K-12–expressing cloned subA and subB resulted in dramatic weight loss. These findings suggest that the toxin may contribute to the pathogenesis of human disease.
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            A new pathogenicity island carrying an allelic variant of the Subtilase cytotoxin is common among Shiga toxin producing Escherichia coli of human and ovine origin.

            Pedro L. Grande, S Morabito, F Scheutz … (2013)
            Subtilase (SubAB) is a cytotoxin elaborated by some Shiga Toxin (Stx)-producing Escherichia coli (STEC) strains usually lacking the locus of enterocyte effacement (LEE). Two variants of SubAB coding genes have been described: subAB(1) , located on the plasmid of the STEC O113 98NK2 strain, and subAB(2) , located on a pathogenicity island (PAI) together with the tia gene, encoding an invasion determinant described in enterotoxigenic E. coli. In the present study, we determined the entire nucleotide sequence of the PAI containing the subAB(2) operon, termed Subtilase-Encoding PAI (SE-PAI), and identified its integration site in the pheV tRNA locus. In addition, a PCR strategy for discriminating the two subAB allelic variants was developed and used to investigate their presence in E. coli strains belonging to different pathotypes and in a large collection of LEE-negative STEC of human and ovine origin. The results confirmed that subAB genes are carried predominantly by STEC and showed their presence in 72% and 86% of the LEE-negative strains from human cases of diarrhoea and from healthy sheep respectively. Most of the subAB-positive strains (98%) identified possessed the subAB(2) allelic variant and were also positive for tia, suggesting the presence of SE-PAI. Altogether, our observations indicate that subAB(2) is the prevalent SubAB-coding operon in LEE-negative STEC circulating in European countries, and that sheep may represent an important reservoir for human infections with these strains. Further studies are needed to assess the role of tia and/or other genes carried by SE-PAI in the colonization of the host intestinal mucosa. © 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.
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              Molecular analysis of subtilase cytotoxin genes of food-borne Shiga toxin-producing Escherichia coli reveals a new allelic subAB variant

              Joschua Funk, Helen Stoeber, Elisabeth Hauser … (2013)
              Background The open reading frames of subAB genes and their flanking regions of 18 food-borne Shiga toxin-producing E. coli (STEC) strains were analyzed. Results All but one subAB open reading frames (ORF) were complete in all STEC strains. The subAB 1 genes of nine STEC strains were located on large plasmids. The subAB 2 allele (here designated subAB 2-1), which was recently described by others to be present in the Subtilase-Encoding PAI (SE-PAI) was found in 6 STEC strains. A new chromosomal subAB 2 variant, designated subAB 2-2 was detected in 6 strains and was linked to a chromosomal gene hypothetically encoding an outer membrane efflux protein (OEP). Three STEC strains contained both subAB 2 variants. DNA analysis indicated sequence conservation in the plasmid-located alleles and sequence heterogeneity among the chromosomal subAB 2 genes. Conclusions The results of this study have shown that 18 subAB-PCR positive STEC strains contain complete subAB open reading frames. Furthermore, the new allelic variant subAB 2-2 was described, which can occur in addition to subAB 2-1 on a new chromosomal locus.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Genome Announc
                Journal ID (iso-abbrev): Genome Announc
                Journal ID (hwp): ga
                Journal ID (pmc): ga
                Journal ID (publisher-id): GA
                Title: Genome Announcements
                Publisher: American Society for Microbiology (1752 N St., N.W., Washington, DC )
                ISSN (Electronic): 2169-8287
                Publication date (Electronic): 23 February 2017
                Publication date Collection: February 2017
                Volume: 5
                Issue: 8
                Electronic Location Identifier: e01582-16
                Affiliations
                [a ]Institute for Food Safety and Hygiene, Vetsuisse Faculty University of Zurich, Zürich, Switzerland
                [b ]Institute of Food, Nutrition and Health, ETH Zürich, Zürich, Switzerland
                [c ]Department of Food Microbiology and Hygiene, Institute of Food Science and Biotechnology, University of Hohenheim, Stuttgart, Germany
                Author notes
                Address correspondence to Roger Stephan, stephanr@ 123456fsafety.uzh.ch .
                Author information
                http://orcid.org/0000-0003-3410-2702
                Article
                Publisher ID: genomeA01582-16
                DOI: 10.1128/genomeA.01582-16
                PMC ID: 5323612
                PubMed ID: 28232433
                SO-VID: ad01a712-bcfd-4311-9a01-7b144872dfd1
                Copyright © Copyright © 2017 Tasara et al.
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

                History
                Date received : 29 November 2016
                Date accepted : 23 December 2016
                Page count
                Figures: 0, Tables: 1, Equations: 0, References: 5, Pages: 2, Words: 1068
                Funding
                Funded by: Swiss Federal Veterinary Office
                Award Recipient : Lisa Fierz
                Funded by: University of Zurich
                Award Recipient : Taurai Tasara
                Categories
                Subject: Prokaryotes
                Custom metadata
                cover-date February 2017

                ScienceOpen disciplines: Genetics
                Data availability:
                ScienceOpen disciplines: Genetics

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