Excision dynamics of <it>Vibrio </it>pathogenicity island-2 from <it>Vibrio cholerae: </it>role of a recombination directionality factor VefA

• Main Authors: Napolitano Michael G, Almagro-Moreno Salvador, Boyd E
• Format: Article
• Language: English
• Published: BMC 2010-11-01
• Series: BMC Microbiology
• Online Access: http://www.biomedcentral.com/1471-2180/10/306

<p>Abstract</p> <p>Background</p> <p><it>Vibrio </it>Pathogenicity Island-2 (VPI-2) is a 57 kb region present in choleragenic <it>V. cholerae </it>isolates that is required for growth on sialic acid as a sole carbon source. <it>V. cholerae </it>non-O1/O139 pathogenic strains also contain VPI-2, which in addition to sialic acid catabolism genes also encodes a type 3 secretion system in these strains. VPI-2 integrates into chromosome 1 at a tRNA-serine site and encodes an integrase <it>intV2 </it>(VC1758) that belongs to the tyrosine recombinase family. IntV2 is required for VPI-2 excision from chromosome 1, which occurs at very low levels, and formation of a non-replicative circular intermediate.</p> <p>Results</p> <p>We determined the conditions and the factors that affect excision of VPI-2 in <it>V. cholerae </it>N16961. We demonstrate that excision from chromosome 1 is induced at low temperature and after sublethal UV-light irradiation treatment. In addition, after UV-light irradiation compared to untreated cells, cells showed increased expression of three genes, <it>intV2 </it>(VC1758)<it/>, and two putative recombination directionality factors (RDFs), <it>vefA </it>(VC1785) and <it>vefB </it>(VC1809) encoded within VPI-2. We demonstrate that along with IntV2, the RDF VefA is essential for excision. We constructed a knockout mutant of <it>vefA </it>in <it>V. cholerae </it>N16961, and found that no excision of VPI-2 occurred, indicating that a functional <it>vefA </it>gene is required for excision. Deletion of the second RDF encoded by <it>vefB </it>did not result in a loss of excision. Among <it>Vibrio </it>species in the genome database, we identified 27 putative RDFs within regions that also encoded IntV2 homologues. Within each species the RDFs and their cognate IntV2 proteins were associated with different island regions suggesting that this pairing is widespread.</p> <p>Conclusions</p> <p>We demonstrate that excision of VPI-2 is induced under some environmental stress conditions and we show for the first time that an RDF encoded within a pathogenicity island in <it>V. cholerae </it>is required for excision of the region.</p>