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Controlling the phosphorylation state of CheY, a response regulator protein, is essential for regulating bacterial chemotaxis and motility. Rapid dephosphorylation of phosphorylated CheY (CheY-P) is crucial for cells to respond to environmental changes. CheY-P dephosphorylation is accomplished by one or more phosphatases in different species, including CheZ, CheC, CheX, FliY, and\/or FliY\/N. Only a<jats:italic>cheX<\/jats:italic>phosphatase homolog has been identified in the<jats:italic>B. burgdorferi<\/jats:italic>genome. However, a role for<jats:italic>cheX<\/jats:italic>in chemotaxis has not been established in any bacterial species. Inactivating<jats:italic>B. burgdorferi cheX<\/jats:italic>by inserting a<jats:italic>flgB-kan<\/jats:italic>cassette resulted in cells (<jats:italic>cheX<\/jats:italic>mutant cells) with a distinct motility phenotype. While wild-type cells ran, paused (stopped or flexed), and reversed, the<jats:italic>cheX<\/jats:italic>mutant cells continuously flexed and were not able to run or reverse. Furthermore, swarm plate and capillary tube chemotaxis assays demonstrated that<jats:italic>cheX<\/jats:italic>mutant cells were deficient in chemotaxis. Wild-type chemotaxis and motility were restored when<jats:italic>cheX<\/jats:italic>mutant cells were complemented with a shuttle vector expressing CheX. Furthermore, CheX dephosphorylated CheY3-P in vitro and eluted as a homodimer in gel filtration chromatography. These findings demonstrated that<jats:italic>B. burgdorferi<\/jats:italic>CheX is a CheY-P phosphatase that is essential for chemotaxis and motility, which is consistent with CheX being the only CheY-P phosphatase in the<jats:italic>B. burgdorferi<\/jats:italic>chemotaxis signal transduction pathway.<\/jats:p>","DOI":"10.1128\/jb.187.23.7963-7969.2005","type":"journal-article","created":{"date-parts":[[2005,11,16]],"date-time":"2005-11-16T02:48:42Z","timestamp":1132109322000},"page":"7963-7969","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":62,"title":["CheX Is a Phosphorylated CheY Phosphatase Essential for<i>Borrelia burgdorferi<\/i>Chemotaxis"],"prefix":"10.1128","volume":"187","author":[{"given":"M. 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