<i>Staphylococcus aureus</i> Proteins Implicated in the Reduced Virulence of <i>sarA</i> and <i>sarA/agr</i> Mutants in Osteomyelitis
Using a murine osteomyelitis model, we recently demonstrated that <i>Staphylococcus aureus sarA</i> and <i>sarA/agr</i> mutants generated in the USA300 strain LAC are attenuated to a greater extent than an isogenic <i>agr</i> mutant and that this can be attributed...
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2025-01-01
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| author | Karen E. Beenken Mara J. Campbell Stephanie D. Byrum Rick D. Edmondson Samuel G. Mackintosh Alan J. Tackett Mark S. Smeltzer |
| author_facet | Karen E. Beenken Mara J. Campbell Stephanie D. Byrum Rick D. Edmondson Samuel G. Mackintosh Alan J. Tackett Mark S. Smeltzer |
| author_sort | Karen E. Beenken |
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| description | Using a murine osteomyelitis model, we recently demonstrated that <i>Staphylococcus aureus sarA</i> and <i>sarA/agr</i> mutants generated in the USA300 strain LAC are attenuated to a greater extent than an isogenic <i>agr</i> mutant and that this can be attributed to a significant extent to the increased production of extracellular proteases in both mutants. Based on this, we used a mass-based proteomics approach to compare the proteomes of LAC, its isogenic <i>agr</i>, <i>sarA</i>, and <i>sarA/agr</i> mutants, and isogenic derivatives of all four of these strains unable to produce the extracellular proteases aureolysin, SspA, SspB, ScpA, or SplA-F. This allowed us to identify proteins that were present in reduced amounts in <i>sarA</i>, and <i>sarA</i>/<i>agr</i> mutants owing to the increased production of extracellular proteases. A total of 1039 proteins were detected in conditioned media (CM) from overnight cultures of LAC, and protease-mediated degradation was shown to contribute to the reduced abundance of 224 of these (21.6%) in CM from the <i>sarA</i> and <i>sarA/agr</i> mutants. Among these were specific proteins previously implicated in the pathogenesis and therapeutic recalcitrance of <i>S. aureus</i> osteomyelitis. This demonstrates that the ability of <i>sarA</i> to limit protease production plays a key role in post-translational remodeling of the <i>S. aureus</i> proteome to a degree that can be correlated with reduced virulence in our osteomyelitis model, and that it does so irrespective of the functional status of <i>agr</i>. This also suggests that at least some of these 224 proteins may be viable targets for prophylactic or therapeutic intervention. |
| format | Article |
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| institution | Kabale University |
| issn | 2076-2607 |
| language | English |
| publishDate | 2025-01-01 |
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| series | Microorganisms |
| spelling | doaj-art-b88c5da26ced4855be0bb8d83f0bd4742025-01-24T13:42:57ZengMDPI AGMicroorganisms2076-26072025-01-0113118110.3390/microorganisms13010181<i>Staphylococcus aureus</i> Proteins Implicated in the Reduced Virulence of <i>sarA</i> and <i>sarA/agr</i> Mutants in OsteomyelitisKaren E. Beenken0Mara J. Campbell1Stephanie D. Byrum2Rick D. Edmondson3Samuel G. Mackintosh4Alan J. Tackett5Mark S. Smeltzer6Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USADepartment of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USADepartment of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USADepartment of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USADepartment of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USADepartment of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USADepartment of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USAUsing a murine osteomyelitis model, we recently demonstrated that <i>Staphylococcus aureus sarA</i> and <i>sarA/agr</i> mutants generated in the USA300 strain LAC are attenuated to a greater extent than an isogenic <i>agr</i> mutant and that this can be attributed to a significant extent to the increased production of extracellular proteases in both mutants. Based on this, we used a mass-based proteomics approach to compare the proteomes of LAC, its isogenic <i>agr</i>, <i>sarA</i>, and <i>sarA/agr</i> mutants, and isogenic derivatives of all four of these strains unable to produce the extracellular proteases aureolysin, SspA, SspB, ScpA, or SplA-F. This allowed us to identify proteins that were present in reduced amounts in <i>sarA</i>, and <i>sarA</i>/<i>agr</i> mutants owing to the increased production of extracellular proteases. A total of 1039 proteins were detected in conditioned media (CM) from overnight cultures of LAC, and protease-mediated degradation was shown to contribute to the reduced abundance of 224 of these (21.6%) in CM from the <i>sarA</i> and <i>sarA/agr</i> mutants. Among these were specific proteins previously implicated in the pathogenesis and therapeutic recalcitrance of <i>S. aureus</i> osteomyelitis. This demonstrates that the ability of <i>sarA</i> to limit protease production plays a key role in post-translational remodeling of the <i>S. aureus</i> proteome to a degree that can be correlated with reduced virulence in our osteomyelitis model, and that it does so irrespective of the functional status of <i>agr</i>. This also suggests that at least some of these 224 proteins may be viable targets for prophylactic or therapeutic intervention.https://www.mdpi.com/2076-2607/13/1/181<i>Staphylococcus</i>osteomyelitis<i>sarA</i><i>agr</i>proteaseproteomics |
| spellingShingle | Karen E. Beenken Mara J. Campbell Stephanie D. Byrum Rick D. Edmondson Samuel G. Mackintosh Alan J. Tackett Mark S. Smeltzer <i>Staphylococcus aureus</i> Proteins Implicated in the Reduced Virulence of <i>sarA</i> and <i>sarA/agr</i> Mutants in Osteomyelitis Microorganisms <i>Staphylococcus</i> osteomyelitis <i>sarA</i> <i>agr</i> protease proteomics |
| title | <i>Staphylococcus aureus</i> Proteins Implicated in the Reduced Virulence of <i>sarA</i> and <i>sarA/agr</i> Mutants in Osteomyelitis |
| title_full | <i>Staphylococcus aureus</i> Proteins Implicated in the Reduced Virulence of <i>sarA</i> and <i>sarA/agr</i> Mutants in Osteomyelitis |
| title_fullStr | <i>Staphylococcus aureus</i> Proteins Implicated in the Reduced Virulence of <i>sarA</i> and <i>sarA/agr</i> Mutants in Osteomyelitis |
| title_full_unstemmed | <i>Staphylococcus aureus</i> Proteins Implicated in the Reduced Virulence of <i>sarA</i> and <i>sarA/agr</i> Mutants in Osteomyelitis |
| title_short | <i>Staphylococcus aureus</i> Proteins Implicated in the Reduced Virulence of <i>sarA</i> and <i>sarA/agr</i> Mutants in Osteomyelitis |
| title_sort | i staphylococcus aureus i proteins implicated in the reduced virulence of i sara i and i sara agr i mutants in osteomyelitis |
| topic | <i>Staphylococcus</i> osteomyelitis <i>sarA</i> <i>agr</i> protease proteomics |
| url | https://www.mdpi.com/2076-2607/13/1/181 |
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