The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrity
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | https://doi.org/10.1038/s41598-019-41461-1 |
Resumo: | This study was funded by the European Research Council through grant ERC-2017-CoG 771709 (to M.G.P.), by Project LISBOA-01-0145-FEDER-007660 Microbiologia Molecular, Estrutural e Celular (to ITQB-NOVA), by the German Research Foundation (DFG; SCHN1284/1-2) to T.S. and FCT fellowship SFRH/BD/71993/2010 (J.M.M.). Mass spectrometry data of muropeptides was acquired by the Mass Spectrometry Unit (UniMS), ITQB/iBET, Oeiras, Portugal. |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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7160 |
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The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrityGeneralThis study was funded by the European Research Council through grant ERC-2017-CoG 771709 (to M.G.P.), by Project LISBOA-01-0145-FEDER-007660 Microbiologia Molecular, Estrutural e Celular (to ITQB-NOVA), by the German Research Foundation (DFG; SCHN1284/1-2) to T.S. and FCT fellowship SFRH/BD/71993/2010 (J.M.M.). Mass spectrometry data of muropeptides was acquired by the Mass Spectrometry Unit (UniMS), ITQB/iBET, Oeiras, Portugal.Bacterial cells are surrounded by cell wall, whose main component is peptidoglycan (PG), a macromolecule that withstands the internal turgor of the cell. PG composition can vary considerably between species. The Gram-positive pathogen Staphylococcus aureus possesses highly crosslinked PG due to the presence of cross bridges containing five glycines, which are synthesised by the FemXAB protein family. FemX adds the first glycine of the cross bridge, while FemA and FemB add the second and the third, and the fourth and the fifth glycines, respectively. Of these, FemX was reported to be essential. To investigate the essentiality of FemAB, we constructed a conditional S. aureus mutant of the femAB operon. Depletion of femAB was lethal, with cells appearing as pseudomulticellular forms that eventually lyse due to extensive membrane rupture. This deleterious effect was mitigated by drastically increasing the osmolarity of the medium, indicating that pentaglycine crosslinks are required for S. aureus cells to withstand internal turgor. Despite the absence of canonical membrane targeting domains, FemA has been shown to localise at the membrane. To study its mechanism of localisation, we constructed mutants in key residues present in the putative transferase pocket and the α6 helix of FemA, possibly involved in tRNA binding. Mutations in the α6 helix led to a sharp decrease in protein activity in vivo and in vitro but did not impair correct membrane localisation, indicating that FemA activity is not required for localisation. Our data indicates that, contrarily to what was previously thought, S. aureus cells do not survive in the absence of a pentaglycine cross bridge.Instituto de Tecnologia Química e Biológica António Xavier (ITQB)DCV - Departamento de Ciências da VidaUCIBIO - Applied Molecular Biosciences UnitMolecular, Structural and Cellular Microbiology (MOSTMICRO)RUNMonteiro, João M.Covas, GonçaloRausch, DanielaFilipe, Sérgio R.Schneider, TanjaSahl, Hans GeorgPinho, Mariana G.2019-07-25T22:33:43Z2019-12-012019-12-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://doi.org/10.1038/s41598-019-41461-1eng2045-2322PURE: 12644811http://www.scopus.com/inward/record.url?scp=85063326118&partnerID=8YFLogxKhttps://doi.org/10.1038/s41598-019-41461-1info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T04:34:51Zoai:run.unl.pt:10362/76533Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:35:38.427635Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrity |
title |
The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrity |
spellingShingle |
The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrity Monteiro, João M. General |
title_short |
The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrity |
title_full |
The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrity |
title_fullStr |
The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrity |
title_full_unstemmed |
The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrity |
title_sort |
The pentaglycine bridges of Staphylococcus aureus peptidoglycan are essential for cell integrity |
author |
Monteiro, João M. |
author_facet |
Monteiro, João M. Covas, Gonçalo Rausch, Daniela Filipe, Sérgio R. Schneider, Tanja Sahl, Hans Georg Pinho, Mariana G. |
author_role |
author |
author2 |
Covas, Gonçalo Rausch, Daniela Filipe, Sérgio R. Schneider, Tanja Sahl, Hans Georg Pinho, Mariana G. |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Instituto de Tecnologia Química e Biológica António Xavier (ITQB) DCV - Departamento de Ciências da Vida UCIBIO - Applied Molecular Biosciences Unit Molecular, Structural and Cellular Microbiology (MOSTMICRO) RUN |
dc.contributor.author.fl_str_mv |
Monteiro, João M. Covas, Gonçalo Rausch, Daniela Filipe, Sérgio R. Schneider, Tanja Sahl, Hans Georg Pinho, Mariana G. |
dc.subject.por.fl_str_mv |
General |
topic |
General |
description |
This study was funded by the European Research Council through grant ERC-2017-CoG 771709 (to M.G.P.), by Project LISBOA-01-0145-FEDER-007660 Microbiologia Molecular, Estrutural e Celular (to ITQB-NOVA), by the German Research Foundation (DFG; SCHN1284/1-2) to T.S. and FCT fellowship SFRH/BD/71993/2010 (J.M.M.). Mass spectrometry data of muropeptides was acquired by the Mass Spectrometry Unit (UniMS), ITQB/iBET, Oeiras, Portugal. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-07-25T22:33:43Z 2019-12-01 2019-12-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://doi.org/10.1038/s41598-019-41461-1 |
url |
https://doi.org/10.1038/s41598-019-41461-1 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
2045-2322 PURE: 12644811 http://www.scopus.com/inward/record.url?scp=85063326118&partnerID=8YFLogxK https://doi.org/10.1038/s41598-019-41461-1 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
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application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799137976941281280 |