title: Comparative analysis of sequences flanking tet(W) resistance genes in multiple species of gut bacteria.
abstract: tet(W) is one of the most abundant tetracycline resistance genes found in bacteria from the mammalian gut and was first identified in the rumen anaerobe Butyrivibrio fibrisolvens 1.230, where it is highly mobile and its transfer is associated with the transposable chromosomal element TnB1230. In order to compare the genetic basis for tet(W) carriage in different bacteria, we studied sequences flanking tet(W) in representatives of seven bacterial genera originating in diverse gut environments. The sequences 657 bp upstream and 43 bp downstream of tet(W) were 96 to 100% similar in all strains examined. A common open reading frame (ORF) was identified downstream of tet(W) in five different bacteria, while another conserved ORF that flanked tet(W) in B. fibrisolvens 1.230 was also present upstream of tet(W) in a human colonic Roseburia isolate and in another rumen B. fibrisolvens isolate. In one species, Bifidobacterium longum (strain F8), a novel transposase was located within the conserved 657-bp region upstream of tet(W) and was flanked by imperfect direct repeats. Additional direct repeats 6 bp long were identified on each end of a chromosomal ORF interrupted by the insertion of the putative transposase and the tet(W) gene. This tet(W) gene was transferable at low frequencies between Bifidobacterium strains. A putative minielement carrying a copy of tet(W) was identified in B. fibrisolvens transconjugants that had acquired the tet(W) gene on TnB1230. Several different mechanisms, including mechanisms involving plasmids and conjugative transposons, appear to be involved in the horizontal transfer of tet(W) genes, but small core regions that may function as minielements are conserved.
authors.affiliation: Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, UK.
authors.forename: Katarzyna A
authors.initials: KA
authors.lastname: Kazimierczak
authors.forename: Harry J
authors.initials: HJ
authors.lastname: Flint
authors.forename: Karen P
authors.initials: KP
authors.lastname: Scott
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000235:genetics* / Q000254:growth & development; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D020134:Catalytic Domain; D003227:Conjugation, Genetic; D017124:Conserved Sequence; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D005798:Genes, Bacterial; D014644:Genetic Variation; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D017422:Sequence Analysis, DNA; D013753:Tetracycline Resistance / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial