YycH and YycI Regulate Expression of <i>Staphylococcus aureus</i> Autolysins by Activation of WalRK Phosphorylation
<i>Staphylococcus aureus</i> is a facultative pathogen that can encode numerous antibiotic resistance and immune evasion genes and can cause severe infections. Reduced susceptibility to last resort antibiotics such as vancomycin and daptomycin is often associated with mutations in <i&...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-06-01
|
| Series: | Microorganisms |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2607/8/6/870 |
| _version_ | 1797565737661890560 |
|---|---|
| author | Mike Gajdiss Ian R. Monk Ute Bertsche Janina Kienemund Tanja Funk Alina Dietrich Michael Hort Esther Sib Timothy P. Stinear Gabriele Bierbaum |
| author_facet | Mike Gajdiss Ian R. Monk Ute Bertsche Janina Kienemund Tanja Funk Alina Dietrich Michael Hort Esther Sib Timothy P. Stinear Gabriele Bierbaum |
| author_sort | Mike Gajdiss |
| collection | DOAJ |
| description | <i>Staphylococcus aureus</i> is a facultative pathogen that can encode numerous antibiotic resistance and immune evasion genes and can cause severe infections. Reduced susceptibility to last resort antibiotics such as vancomycin and daptomycin is often associated with mutations in <i>walRK</i>, an essential two-component regulatory system (TCS). This study focuses on the WalK accessory membrane proteins YycH and YycI and their influence on WalRK phosphorylation. Depletion of YycH and YycI by antisense RNA caused an impaired autolysis, indicating a positive regulatory function on WalK as has been previously described. Phosphorylation assays with full-length recombinant proteins in phospholipid liposomes showed that YycH and YycI stimulate WalK activity and that both regulatory proteins are needed for full activation of the WalK kinase. This was validated in vivo through examining the phosphorylation status of WalR using Phos-tag SDS-PAGE with a <i>yycHI</i> deletion mutant exhibiting reduced levels of phosphorylated WalR. In the <i>yycHI</i> knockdown strain, muropeptide composition of the cell wall was not affected, however, the wall teichoic acid content was increased. In conclusion, a direct modulation of WalRK phosphorylation activity by the accessory proteins YycH and YycI is reported both in vitro and in vivo. Taken together, our results show that YycH and YycI are important in the direct regulation of WalRK-dependent cell wall metabolism. |
| first_indexed | 2024-03-10T19:17:17Z |
| format | Article |
| id | doaj.art-c2b6105d08294a348f125df64b053a65 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T19:17:17Z |
| publishDate | 2020-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-c2b6105d08294a348f125df64b053a652023-11-20T03:17:37ZengMDPI AGMicroorganisms2076-26072020-06-018687010.3390/microorganisms8060870YycH and YycI Regulate Expression of <i>Staphylococcus aureus</i> Autolysins by Activation of WalRK PhosphorylationMike Gajdiss0Ian R. Monk1Ute Bertsche2Janina Kienemund3Tanja Funk4Alina Dietrich5Michael Hort6Esther Sib7Timothy P. Stinear8Gabriele Bierbaum9Institute of Medical Microbiology, Immunology and Parasitology, University Hospital Bonn, 53125 Bonn, GermanyDepartment of Microbiology and Immunology, Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC 3010, AustraliaDepartment of Infection Biology, University of Tuebingen, 72076 Tuebingen, GermanyInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital Bonn, 53125 Bonn, GermanyInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital Bonn, 53125 Bonn, GermanyInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital Bonn, 53125 Bonn, GermanyInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital Bonn, 53125 Bonn, GermanyInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital Bonn, 53125 Bonn, GermanyDepartment of Microbiology and Immunology, Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC 3010, AustraliaInstitute of Medical Microbiology, Immunology and Parasitology, University Hospital Bonn, 53125 Bonn, Germany<i>Staphylococcus aureus</i> is a facultative pathogen that can encode numerous antibiotic resistance and immune evasion genes and can cause severe infections. Reduced susceptibility to last resort antibiotics such as vancomycin and daptomycin is often associated with mutations in <i>walRK</i>, an essential two-component regulatory system (TCS). This study focuses on the WalK accessory membrane proteins YycH and YycI and their influence on WalRK phosphorylation. Depletion of YycH and YycI by antisense RNA caused an impaired autolysis, indicating a positive regulatory function on WalK as has been previously described. Phosphorylation assays with full-length recombinant proteins in phospholipid liposomes showed that YycH and YycI stimulate WalK activity and that both regulatory proteins are needed for full activation of the WalK kinase. This was validated in vivo through examining the phosphorylation status of WalR using Phos-tag SDS-PAGE with a <i>yycHI</i> deletion mutant exhibiting reduced levels of phosphorylated WalR. In the <i>yycHI</i> knockdown strain, muropeptide composition of the cell wall was not affected, however, the wall teichoic acid content was increased. In conclusion, a direct modulation of WalRK phosphorylation activity by the accessory proteins YycH and YycI is reported both in vitro and in vivo. Taken together, our results show that YycH and YycI are important in the direct regulation of WalRK-dependent cell wall metabolism.https://www.mdpi.com/2076-2607/8/6/870WalKWalRtwo-component regulatory system<i>Staphylococcus aureus</i>YycHYycI |
| spellingShingle | Mike Gajdiss Ian R. Monk Ute Bertsche Janina Kienemund Tanja Funk Alina Dietrich Michael Hort Esther Sib Timothy P. Stinear Gabriele Bierbaum YycH and YycI Regulate Expression of <i>Staphylococcus aureus</i> Autolysins by Activation of WalRK Phosphorylation Microorganisms WalK WalR two-component regulatory system <i>Staphylococcus aureus</i> YycH YycI |
| title | YycH and YycI Regulate Expression of <i>Staphylococcus aureus</i> Autolysins by Activation of WalRK Phosphorylation |
| title_full | YycH and YycI Regulate Expression of <i>Staphylococcus aureus</i> Autolysins by Activation of WalRK Phosphorylation |
| title_fullStr | YycH and YycI Regulate Expression of <i>Staphylococcus aureus</i> Autolysins by Activation of WalRK Phosphorylation |
| title_full_unstemmed | YycH and YycI Regulate Expression of <i>Staphylococcus aureus</i> Autolysins by Activation of WalRK Phosphorylation |
| title_short | YycH and YycI Regulate Expression of <i>Staphylococcus aureus</i> Autolysins by Activation of WalRK Phosphorylation |
| title_sort | yych and yyci regulate expression of i staphylococcus aureus i autolysins by activation of walrk phosphorylation |
| topic | WalK WalR two-component regulatory system <i>Staphylococcus aureus</i> YycH YycI |
| url | https://www.mdpi.com/2076-2607/8/6/870 |
| work_keys_str_mv | AT mikegajdiss yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation AT ianrmonk yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation AT utebertsche yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation AT janinakienemund yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation AT tanjafunk yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation AT alinadietrich yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation AT michaelhort yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation AT esthersib yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation AT timothypstinear yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation AT gabrielebierbaum yychandyyciregulateexpressionofistaphylococcusaureusiautolysinsbyactivationofwalrkphosphorylation |