Cargando…

YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation

Staphylococcus aureus 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 walRK, an essential two-c...

Descripción completa

Detalles Bibliográficos
Autores principales: Gajdiss, Mike, Monk, Ian R., Bertsche, Ute, Kienemund, Janina, Funk, Tanja, Dietrich, Alina, Hort, Michael, Sib, Esther, Stinear, Timothy P., Bierbaum, Gabriele
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7355866/
https://www.ncbi.nlm.nih.gov/pubmed/32526915
http://dx.doi.org/10.3390/microorganisms8060870
_version_ 1783558374470713344
author Gajdiss, Mike
Monk, Ian R.
Bertsche, Ute
Kienemund, Janina
Funk, Tanja
Dietrich, Alina
Hort, Michael
Sib, Esther
Stinear, Timothy P.
Bierbaum, Gabriele
author_facet Gajdiss, Mike
Monk, Ian R.
Bertsche, Ute
Kienemund, Janina
Funk, Tanja
Dietrich, Alina
Hort, Michael
Sib, Esther
Stinear, Timothy P.
Bierbaum, Gabriele
author_sort Gajdiss, Mike
collection PubMed
description Staphylococcus aureus 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 walRK, 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 yycHI deletion mutant exhibiting reduced levels of phosphorylated WalR. In the yycHI 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.
format Online
Article
Text
id pubmed-7355866
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-73558662020-07-23 YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation Gajdiss, Mike Monk, Ian R. Bertsche, Ute Kienemund, Janina Funk, Tanja Dietrich, Alina Hort, Michael Sib, Esther Stinear, Timothy P. Bierbaum, Gabriele Microorganisms Article Staphylococcus aureus 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 walRK, 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 yycHI deletion mutant exhibiting reduced levels of phosphorylated WalR. In the yycHI 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. MDPI 2020-06-09 /pmc/articles/PMC7355866/ /pubmed/32526915 http://dx.doi.org/10.3390/microorganisms8060870 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gajdiss, Mike
Monk, Ian R.
Bertsche, Ute
Kienemund, Janina
Funk, Tanja
Dietrich, Alina
Hort, Michael
Sib, Esther
Stinear, Timothy P.
Bierbaum, Gabriele
YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation
title YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation
title_full YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation
title_fullStr YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation
title_full_unstemmed YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation
title_short YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation
title_sort yych and yyci regulate expression of staphylococcus aureus autolysins by activation of walrk phosphorylation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7355866/
https://www.ncbi.nlm.nih.gov/pubmed/32526915
http://dx.doi.org/10.3390/microorganisms8060870
work_keys_str_mv AT gajdissmike yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation
AT monkianr yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation
AT bertscheute yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation
AT kienemundjanina yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation
AT funktanja yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation
AT dietrichalina yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation
AT hortmichael yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation
AT sibesther yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation
AT stineartimothyp yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation
AT bierbaumgabriele yychandyyciregulateexpressionofstaphylococcusaureusautolysinsbyactivationofwalrkphosphorylation