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In this study, we have employed the intrinsically resistant opportunistic pathogen <jats:italic>Stenotrophomonas maltophilia<\/jats:italic> as a model to study the mechanisms involved in the acquisition of mutation-driven resistance to antibiotics. To this aim, laboratory experimental evolution studies, followed by whole-genome sequencing, were performed in the presence of the third-generation cephalosporin ceftazidime. Using this approach, we determined that exposure to increasing concentrations of ceftazidime selects high-level resistance in <jats:italic>S. maltophilia<\/jats:italic> through a novel mechanism: amino acid substitutions in SmeH, the transporter protein of the SmeGH RND efflux pump. The recreation of these mutants in a wild-type background demonstrated that, in addition to ceftazidime, the existence of these substitutions provides bacteria with cross-resistance to other beta-lactam drugs. This acquired resistance does not impose relevant fitness costs when bacteria grow in the absence of antibiotics. Structural prediction of both amino acid residues points that the observed resistance phenotype could be driven by changes in substrate access and recognition.<\/jats:p>","DOI":"10.1038\/s41598-019-41308-9","type":"journal-article","created":{"date-parts":[[2019,3,20]],"date-time":"2019-03-20T11:03:45Z","timestamp":1553079825000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":40,"title":["Involvement of the RND efflux pump transporter SmeH in the acquisition of resistance to ceftazidime in Stenotrophomonas maltophilia"],"prefix":"10.1038","volume":"9","author":[{"given":"Paula","family":"Blanco","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando","family":"Corona","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8813-7607","authenticated-orcid":false,"given":"Jos\u00e9 Luis","family":"Mart\u00ednez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,3,20]]},"reference":[{"key":"41308_CR1","doi-asserted-by":"publisher","first-page":"949","DOI":"10.1002\/phar.1636","volume":"35","author":"KS Kaye","year":"2015","unstructured":"Kaye, K. 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