Site-specific integration of the conjugal Vibrio cholerae SXT element into prfC.
Vibrio cholerae O139, the first non-O1 serogroup of V. cholerae to give rise to epidemic cholera, is characteristically resistant to the antibiotics sulphamethoxazole, trimethoprim, chloramphenicol and streptomycin. Resistances to these antibiotics are encoded by a 62 kb self-transmissible, conjugative, chromosomally integrating element designated the 'SXT element'. We found that the SXT element integrates site specifically into both V. cholerae and Escherichia coli K-12 into the 5' end of prfC, the gene encoding peptide chain release factor 3. Integration of the SXT element interrupts the chromosomal prfC gene, but the element encodes a new 5' end of prfC that restores the reading frame of this gene. The recombinant of prfC allele created upon element integration is functional. The integration and excision mechanism of the SXT element shares many features with site-specific recombination found in lambdoid phages. First, like lambda, the SXT element forms a circular extrachromosomal intermediate through specific recombination of the left and right ends of the integrated element. Second, chromosomal integration of the element occurs via site-specific recombination in a 17 bp sequence found in the circular form of the SXT element and a similar 17 bp sequence in prfC. Third, both chromosomal integration and excision of the SXT element were found to require an element-encoded int gene with strong similarities to the lambda integrase family. Based on the properties of the SXT element, we propose to classify this element as a CONSTIN, an acronym for a conjugative, self-transmissible, integrating element.
Hochhut
B
B
New England Medical Center, Boston, MA, USA.
Waldor
M K
MK
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003227:Conjugation, Genetic / Q000235:genetics; D004251:DNA Transposable Elements; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics; D019426:Integrases / Q000235:genetics*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010454:Peptide Termination Factors / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D014734:Vibrio cholerae / Q000235:genetics* / Q000502:physiology*; D001616:beta-Galactosidase / Q000378:metabolism
D004251:DNA Transposable Elements; D010454:Peptide Termination Factors; C087971:peptide-chain-release factor 3; D019426:Integrases; D001616:beta-Galactosidase