title: Stability of the Encoding Plasmids and Surface Expression of CS6 Differs in Enterotoxigenic Escherichia coli (ETEC) Encoding Different Heat-Stable (ST) Enterotoxins (STh and STp).
abstract: Enterotoxigenic Escherichia coli (ETEC), one of the most common reasons of diarrhea among infants and children in developing countries, causes disease by expression of either or both of the enterotoxins heat-labile (LT) and heat-stable (ST; divided into human-type [STh] and porcine-type [STp] variants), and colonization factors (CFs) among which CS6 is one of the most prevalent ETEC CFs. In this study we show that ETEC isolates expressing CS6+STh have higher copy numbers of the cssABCD operon encoding CS6 than those expressing CS6+STp. Long term cultivation of up to ten over-night passages of ETEC isolates harboring CS6+STh (n = 10) or CS6+STp (n = 15) showed instability of phenotypic expression of CS6 in a majority of the CS6+STp isolates, whereas most of the CS6+STh isolates retained CS6 expression. The observed instability was a correlated with loss of genes cssA and cssD as examined by PCR. Mobilization of the CS6 plasmid from an unstable CS6+STp isolate into a laboratory E. coli strain resulted in loss of the plasmid after a single over-night passage whereas the plasmid from an CS6+STh strain was retained in the laboratory strain during 10 passages. A sequence comparison between the CS6 plasmids from a stable and an unstable ETEC isolate revealed that genes necessary for plasmid stabilization, for example pemI, pemK, stbA, stbB and parM, were not present in the unstable ETEC isolate. Our results indicate that stable retention of CS6 may in part be affected by the stability of the plasmid on which both CS6 and STp or STh are located.
authors.affiliation: Department of Microbiology and Immunology, Institute of Biomedicine, University of Gothenburg, S-40530, Göteborg, Sweden.
authors.forename: Joshua
authors.initials: J
authors.lastname: Tobias
authors.affiliation: Department of Microbiology and Immunology, Institute of Biomedicine, University of Gothenburg, S-40530, Göteborg, Sweden.
authors.forename: Astrid
authors.initials: A
authors.lastname: Von Mentzer
authors.affiliation: Department of Microbiology and Immunology, Institute of Biomedicine, University of Gothenburg, S-40530, Göteborg, Sweden.
authors.forename: Patricia
authors.initials: P
authors.lastname: Loayza Frykberg
authors.affiliation: Pathogen Genomics, Wellcome Trust Sanger Institute, Hinxton, Cambridge, CB10 1SA, United Kingdom.
authors.forename: Martin
authors.initials: M
authors.lastname: Aslett
authors.affiliation: Pathogen Genomics, Wellcome Trust Sanger Institute, Hinxton, Cambridge, CB10 1SA, United Kingdom.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Page
authors.affiliation: Department of Microbiology and Immunology, Institute of Biomedicine, University of Gothenburg, S-40530, Göteborg, Sweden.|Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, S-171 77, Stockholm, Sweden.
authors.forename: Åsa
authors.initials: Å
authors.lastname: Sjöling
authors.affiliation: Department of Microbiology and Immunology, Institute of Biomedicine, University of Gothenburg, S-40530, Göteborg, Sweden.
authors.forename: Ann-Mari
authors.initials: AM
authors.lastname: Svennerholm
mesh_terms: D054307:Enterotoxigenic Escherichia coli / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000472:pathogenicity; D004768:Enterotoxins / Q000737:chemistry / Q000378:metabolism*; D004927:Escherichia coli Infections / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D009154:Mutation / Q000235:genetics; D009876:Operon; D010957:Plasmids / Q000737:chemistry* / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D004768:Enterotoxins; D029968:Escherichia coli Proteins