Role of <i>dipA</i> and <i>pilD</i> in <i>Francisella tularensis</i> Susceptibility to Resazurin
The phenoxazine dye resazurin exhibits bactericidal activity against the Gram-negative pathogens <i>Francisella tularensis</i> and <i>Neisseria gonorrhoeae</i>. One resazurin derivative, resorufin pentyl ether, significantly reduces vaginal colonization by <i>Neisseria...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-08-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/10/8/992 |
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| author | Kendall Souder Emma J. Beatty Siena C. McGovern Michael Whaby Emily Young Jacob Pancake Daron Weekley Justin Rice Donald A. Primerano James Denvir Joseph Horzempa Deanna M. Schmitt |
| author_facet | Kendall Souder Emma J. Beatty Siena C. McGovern Michael Whaby Emily Young Jacob Pancake Daron Weekley Justin Rice Donald A. Primerano James Denvir Joseph Horzempa Deanna M. Schmitt |
| author_sort | Kendall Souder |
| collection | DOAJ |
| description | The phenoxazine dye resazurin exhibits bactericidal activity against the Gram-negative pathogens <i>Francisella tularensis</i> and <i>Neisseria gonorrhoeae</i>. One resazurin derivative, resorufin pentyl ether, significantly reduces vaginal colonization by <i>Neisseria gonorrhoeae</i> in a mouse model of infection. The narrow spectrum of bacteria susceptible to resazurin and its derivatives suggests these compounds have a novel mode of action. To identify potential targets of resazurin and mechanisms of resistance, we isolated mutants of <i>F. tularensis</i> subsp. <i>holarctica</i> live vaccine strain (LVS) exhibiting reduced susceptibility to resazurin and performed whole genome sequencing. The genes <i>pilD</i> (FTL_0959) and <i>dipA</i> (FTL_1306) were mutated in half of the 46 resazurin-resistant (RZR) strains sequenced. Complementation of select RZR LVS isolates with wild-type <i>dipA</i> or <i>pilD</i> partially restored sensitivity to resazurin. To further characterize the role of <i>dipA</i> and <i>pilD</i> in resazurin susceptibility, a <i>dipA</i> deletion mutant, Δ<i>dipA</i>, and <i>pilD</i> disruption mutant, FTL_0959d, were generated. Both mutants were less sensitive to killing by resazurin compared to wild-type LVS with phenotypes similar to the spontaneous resazurin-resistant mutants. This study identified a novel role for two genes <i>dipA</i> and <i>pilD</i> in <i>F. tularensis</i> susceptibility to resazurin. |
| first_indexed | 2024-03-10T09:03:25Z |
| format | Article |
| id | doaj.art-842dbe0aa9054170a171bc95e0ed99c4 |
| institution | Directory Open Access Journal |
| issn | 2079-6382 |
| language | English |
| last_indexed | 2024-03-10T09:03:25Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibiotics |
| spelling | doaj.art-842dbe0aa9054170a171bc95e0ed99c42023-11-22T06:34:22ZengMDPI AGAntibiotics2079-63822021-08-0110899210.3390/antibiotics10080992Role of <i>dipA</i> and <i>pilD</i> in <i>Francisella tularensis</i> Susceptibility to ResazurinKendall Souder0Emma J. Beatty1Siena C. McGovern2Michael Whaby3Emily Young4Jacob Pancake5Daron Weekley6Justin Rice7Donald A. Primerano8James Denvir9Joseph Horzempa10Deanna M. Schmitt11Department of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USADepartment of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USADepartment of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USADepartment of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USADepartment of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USADepartment of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USADepartment of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USADepartment of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USADepartment of Biomedical Sciences, Joan C. Edwards School of Medicine, Marshall University, Huntington, WV 25755, USADepartment of Biomedical Sciences, Joan C. Edwards School of Medicine, Marshall University, Huntington, WV 25755, USADepartment of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USADepartment of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USAThe phenoxazine dye resazurin exhibits bactericidal activity against the Gram-negative pathogens <i>Francisella tularensis</i> and <i>Neisseria gonorrhoeae</i>. One resazurin derivative, resorufin pentyl ether, significantly reduces vaginal colonization by <i>Neisseria gonorrhoeae</i> in a mouse model of infection. The narrow spectrum of bacteria susceptible to resazurin and its derivatives suggests these compounds have a novel mode of action. To identify potential targets of resazurin and mechanisms of resistance, we isolated mutants of <i>F. tularensis</i> subsp. <i>holarctica</i> live vaccine strain (LVS) exhibiting reduced susceptibility to resazurin and performed whole genome sequencing. The genes <i>pilD</i> (FTL_0959) and <i>dipA</i> (FTL_1306) were mutated in half of the 46 resazurin-resistant (RZR) strains sequenced. Complementation of select RZR LVS isolates with wild-type <i>dipA</i> or <i>pilD</i> partially restored sensitivity to resazurin. To further characterize the role of <i>dipA</i> and <i>pilD</i> in resazurin susceptibility, a <i>dipA</i> deletion mutant, Δ<i>dipA</i>, and <i>pilD</i> disruption mutant, FTL_0959d, were generated. Both mutants were less sensitive to killing by resazurin compared to wild-type LVS with phenotypes similar to the spontaneous resazurin-resistant mutants. This study identified a novel role for two genes <i>dipA</i> and <i>pilD</i> in <i>F. tularensis</i> susceptibility to resazurin.https://www.mdpi.com/2079-6382/10/8/992<i>Francisella tularensis</i>resazurinDipAPilDtularemiaantimicrobial |
| spellingShingle | Kendall Souder Emma J. Beatty Siena C. McGovern Michael Whaby Emily Young Jacob Pancake Daron Weekley Justin Rice Donald A. Primerano James Denvir Joseph Horzempa Deanna M. Schmitt Role of <i>dipA</i> and <i>pilD</i> in <i>Francisella tularensis</i> Susceptibility to Resazurin Antibiotics <i>Francisella tularensis</i> resazurin DipA PilD tularemia antimicrobial |
| title | Role of <i>dipA</i> and <i>pilD</i> in <i>Francisella tularensis</i> Susceptibility to Resazurin |
| title_full | Role of <i>dipA</i> and <i>pilD</i> in <i>Francisella tularensis</i> Susceptibility to Resazurin |
| title_fullStr | Role of <i>dipA</i> and <i>pilD</i> in <i>Francisella tularensis</i> Susceptibility to Resazurin |
| title_full_unstemmed | Role of <i>dipA</i> and <i>pilD</i> in <i>Francisella tularensis</i> Susceptibility to Resazurin |
| title_short | Role of <i>dipA</i> and <i>pilD</i> in <i>Francisella tularensis</i> Susceptibility to Resazurin |
| title_sort | role of i dipa i and i pild i in i francisella tularensis i susceptibility to resazurin |
| topic | <i>Francisella tularensis</i> resazurin DipA PilD tularemia antimicrobial |
| url | https://www.mdpi.com/2079-6382/10/8/992 |
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