Effect of a Point Mutation in mprF on Susceptibility to Daptomycin, Vancomycin, and Oxacillin in an MRSA Clinical Strain
We previously reported the sequential recovery of daptomycin-nonsusceptible MRSA clinical isolates with an L431F substitution in the MprF protein. The aim of the present study is to determine the effect of this mutation by replacing the mprF gene on the chromosome of a daptomycin-susceptible progeni...
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Frontiers Media S.A.
2018-05-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2018.01086/full |
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| author | Feng-Jui Chen Tsai-Ling Lauderdale Chen-Hsiang Lee Yu-Chieh Hsu I-Wen Huang Pei-Chi Hsu Chung-Shi Yang |
| author_facet | Feng-Jui Chen Tsai-Ling Lauderdale Chen-Hsiang Lee Yu-Chieh Hsu I-Wen Huang Pei-Chi Hsu Chung-Shi Yang |
| author_sort | Feng-Jui Chen |
| collection | DOAJ |
| description | We previously reported the sequential recovery of daptomycin-nonsusceptible MRSA clinical isolates with an L431F substitution in the MprF protein. The aim of the present study is to determine the effect of this mutation by replacing the mprF gene on the chromosome of a daptomycin-susceptible progenitor strain, CGK5, to obtain CGK5mut having the L431F MprF mutation. Compared to CGK5, the daptomycin and vancomycin MICs of CGK5mut increased from 0.5 to 3 μg/ml and from 1.5 to 3 μg/ml, respectively; however, its oxacillin MIC decreased from 128 to 1 μg/ml in medium without added 2% NaCl. The expression levels of vraSR and several other cell-wall synthesis-related genes were significantly increased in CGK5mut, and the mutant also had significantly reduced negative cell membrane charge, thicker cell wall, and longer doubling time. These features were abolished in the reverse mutant carrying F431L MprF, confirming the pleiotropic effects of the L431F MprF mutation. We believe that this is the first work that shows a single MprF missense mutation can lead to not only changes in the cell membrane but also increased expression of vraSR and subsequently increased resistance to daptomycin and vancomycin while simultaneously conferring increased susceptibility to oxacillin in an isogenic MRSA strain. |
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| last_indexed | 2024-12-21T22:38:56Z |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-4bbe3309f5244af2abe683a587c74cd02022-12-21T18:47:53ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-05-01910.3389/fmicb.2018.01086369299Effect of a Point Mutation in mprF on Susceptibility to Daptomycin, Vancomycin, and Oxacillin in an MRSA Clinical StrainFeng-Jui Chen0Tsai-Ling Lauderdale1Chen-Hsiang Lee2Yu-Chieh Hsu3I-Wen Huang4Pei-Chi Hsu5Chung-Shi Yang6National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, TaiwanNational Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, TaiwanDivision of Infectious Diseases, Kaohsiung Chang Gung Memorial Hospital, College of Medicine, Chang Gung University, Kaohsiung, TaiwanNational Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, TaiwanNational Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, TaiwanNational Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, TaiwanInstitute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Zhunan, TaiwanWe previously reported the sequential recovery of daptomycin-nonsusceptible MRSA clinical isolates with an L431F substitution in the MprF protein. The aim of the present study is to determine the effect of this mutation by replacing the mprF gene on the chromosome of a daptomycin-susceptible progenitor strain, CGK5, to obtain CGK5mut having the L431F MprF mutation. Compared to CGK5, the daptomycin and vancomycin MICs of CGK5mut increased from 0.5 to 3 μg/ml and from 1.5 to 3 μg/ml, respectively; however, its oxacillin MIC decreased from 128 to 1 μg/ml in medium without added 2% NaCl. The expression levels of vraSR and several other cell-wall synthesis-related genes were significantly increased in CGK5mut, and the mutant also had significantly reduced negative cell membrane charge, thicker cell wall, and longer doubling time. These features were abolished in the reverse mutant carrying F431L MprF, confirming the pleiotropic effects of the L431F MprF mutation. We believe that this is the first work that shows a single MprF missense mutation can lead to not only changes in the cell membrane but also increased expression of vraSR and subsequently increased resistance to daptomycin and vancomycin while simultaneously conferring increased susceptibility to oxacillin in an isogenic MRSA strain.https://www.frontiersin.org/article/10.3389/fmicb.2018.01086/fullMRSAevolutiondrug resistancedaptomycinvancomycinoxacillin |
| spellingShingle | Feng-Jui Chen Tsai-Ling Lauderdale Chen-Hsiang Lee Yu-Chieh Hsu I-Wen Huang Pei-Chi Hsu Chung-Shi Yang Effect of a Point Mutation in mprF on Susceptibility to Daptomycin, Vancomycin, and Oxacillin in an MRSA Clinical Strain Frontiers in Microbiology MRSA evolution drug resistance daptomycin vancomycin oxacillin |
| title | Effect of a Point Mutation in mprF on Susceptibility to Daptomycin, Vancomycin, and Oxacillin in an MRSA Clinical Strain |
| title_full | Effect of a Point Mutation in mprF on Susceptibility to Daptomycin, Vancomycin, and Oxacillin in an MRSA Clinical Strain |
| title_fullStr | Effect of a Point Mutation in mprF on Susceptibility to Daptomycin, Vancomycin, and Oxacillin in an MRSA Clinical Strain |
| title_full_unstemmed | Effect of a Point Mutation in mprF on Susceptibility to Daptomycin, Vancomycin, and Oxacillin in an MRSA Clinical Strain |
| title_short | Effect of a Point Mutation in mprF on Susceptibility to Daptomycin, Vancomycin, and Oxacillin in an MRSA Clinical Strain |
| title_sort | effect of a point mutation in mprf on susceptibility to daptomycin vancomycin and oxacillin in an mrsa clinical strain |
| topic | MRSA evolution drug resistance daptomycin vancomycin oxacillin |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2018.01086/full |
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