{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T08:26:24Z","timestamp":1791275184419,"version":"4.1.0"},"reference-count":46,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2014,3,14]],"date-time":"2014-03-14T00:00:00Z","timestamp":1394755200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2014,4]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\"><jats:italic>V<\/jats:italic><\/jats:styled-content><jats:italic>ibrio cholerae<\/jats:italic>, the causative agent of cholera, remains a threat to public health in areas with inadequate sanitation. As a waterborne pathogen, <jats:styled-content style=\"fixed-case\"><jats:italic>V<\/jats:italic><\/jats:styled-content><jats:italic>.\u2009cholerae<\/jats:italic> moves between two dissimilar environments, aquatic reservoirs and the intestinal tract of humans. Accordingly, this pathogen undergoes adaptive shifts in gene expression throughout the different stages of its lifecycle. One particular gene, <jats:styled-content style=\"fixed-case\"><jats:italic>xds<\/jats:italic><\/jats:styled-content>, encodes a secreted exonuclease that was previously identified as being induced during infection. Here we sought to identify regulators responsible for the <jats:italic>in vivo<\/jats:italic>\u2010specific induction of <jats:styled-content style=\"fixed-case\"><jats:italic>xds<\/jats:italic><\/jats:styled-content>. A transcriptional fusion of <jats:styled-content style=\"fixed-case\"><jats:italic>xds<\/jats:italic><\/jats:styled-content> to two consecutive antibiotic resistance genes was used to select transposon mutants that had inserted within or adjacent to regulatory genes and thereby caused increased expression of the <jats:styled-content style=\"fixed-case\"><jats:italic>xds<\/jats:italic><\/jats:styled-content> fusion under non\u2010inducing conditions. Large pools of selected insertion sites were sequenced in a high throughput manner using Tn\u2010seq to identify potential mechanisms of <jats:styled-content style=\"fixed-case\"><jats:italic>xds<\/jats:italic><\/jats:styled-content> regulation. Our selection identified the two\u2010component system <jats:styled-content style=\"fixed-case\">PhoB<\/jats:styled-content>\/<jats:styled-content style=\"fixed-case\">R<\/jats:styled-content> as the dominant activator of <jats:styled-content style=\"fixed-case\"><jats:italic>xds<\/jats:italic><\/jats:styled-content> expression. <jats:italic>In vitro<\/jats:italic> validation confirmed that <jats:styled-content style=\"fixed-case\">PhoB<\/jats:styled-content>, a protein which is only active during phosphate limitation, was responsible for <jats:styled-content style=\"fixed-case\"><jats:italic>xds<\/jats:italic><\/jats:styled-content> activation. Using <jats:styled-content style=\"fixed-case\"><jats:italic>xds<\/jats:italic><\/jats:styled-content> expression as a biosensor of the extracellular phosphate level, we observed that the mouse small intestine is a phosphate\u2010limited environment.<\/jats:p>","DOI":"10.1111\/mmi.12557","type":"journal-article","created":{"date-parts":[[2014,2,20]],"date-time":"2014-02-20T17:17:22Z","timestamp":1392916642000},"page":"302-315","source":"Crossref","is-referenced-by-count":41,"title":["Identification of <i>in vivo<\/i> regulators of the <scp><i>V<\/i><\/scp><i>ibrio cholerae<\/i>\u2005<scp><i>xds<\/i><\/scp> gene using a high\u2010throughput genetic selection"],"prefix":"10.1111","volume":"92","author":[{"given":"EmilyKate","family":"McDonough","sequence":"first","affiliation":[{"name":"Howard Hughes Medical Institute and Department of Molecular Biology and Microbiology Tufts University School of Medicine  Boston MA 02111 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