{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T14:37:42Z","timestamp":1792075062521,"version":"4.4.0"},"reference-count":52,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,10,24]],"date-time":"2024-10-24T00:00:00Z","timestamp":1729728000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Daptomycin non-susceptible <jats:italic>Staphylococcus aureus<\/jats:italic> (DNS) strains pose a serious clinical threat, yet their characteristics remain poorly understood.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>DNS derivatives were generated by exposing <jats:italic>S. aureus<\/jats:italic> strains to subinhibitory concentrations of daptomycin. Competition experiment and growth kinetics experiment were used to observe the growth of bacteria. <jats:italic>Galleria mellonella<\/jats:italic> larvae and mouse skin abscess models were used to observe the virulence of bacteria. Transmission electron microscopy (TEM), cytochrome C experiment and biofilm formation experiment were used to observe the drug resistance phenotype. And homologous recombination was used to study the role of mutations.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Phenotypic profiling of DNS strains revealed impaired growth, increased cell wall thickness, enhanced biofilm formation, reduced negative surface charge, and attenuated virulence compared to their wild-type strains. Whole genome sequencing identified mutations in <jats:italic>mprF<\/jats:italic>, <jats:italic>cls2<\/jats:italic>, and <jats:italic>saeR<\/jats:italic> in DNS strains. Allelic replacement experiments validated the roles of MprF L341F and Cls2 F60S substitutions in augmenting daptomycin non-susceptibility in Newman. Deletion of <jats:italic>saeR<\/jats:italic> in the Newman<jats:sub>MprFL341F<\/jats:sub> strain and complementation of <jats:italic>saeR<\/jats:italic> in the Newman-DNS strain did not directly alter daptomycin susceptibility. However, the deletion of <jats:italic>saeR<\/jats:italic> was found to enhance competitive fitness under daptomycin pressure.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This work validates adaptive laboratory evolution (ALE) for modeling clinical DNS strains and uncovers contributions of <jats:italic>mprF<\/jats:italic>, <jats:italic>cls2<\/jats:italic>, and <jats:italic>saeR<\/jats:italic> mutations to the adaptation and resistance mechanisms of <jats:italic>S. aureus<\/jats:italic> against daptomycin. These findings enrich our understanding of how <jats:italic>S. aureus<\/jats:italic> acquired resistance to daptomycin, thus paving the way for the development of more effective treatment strategies and offering potential molecular markers for resistance surveillance.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2024.1453233","type":"journal-article","created":{"date-parts":[[2024,10,24]],"date-time":"2024-10-24T05:10:21Z","timestamp":1729746621000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Phenotypic and genetic characterization of daptomycin non-susceptible Staphylococcus aureus strains selected by adaptive laboratory evolution"],"prefix":"10.3389","volume":"14","author":[{"given":"Yanlei","family":"Xu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanghua","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huilin","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bingjie","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingyi","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongpeng","family":"Shang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaocui","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinjuan","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fangyou","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,10,24]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkp155","article-title":"Comparison of the pharmacokinetics, safety and tolerability of daptomycin in healthy adult volunteers following intravenous administration by 30 min infusion or 2 min injection","volume":"64","author":"Abhijit","year":"2009","journal-title":"J.\u00a0Antimicrob. 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