Cargando…

Structural basis of broad-spectrum β-lactam resistance in Staphylococcus aureus

Broad-spectrum β-lactam antibiotic resistance in Staphylococcus aureus is a global healthcare burden(1,2). In clinical strains, resistance is largely controlled by BlaR1(3), a receptor that senses β-lactams through the acylation of its sensor domain, inducing transmembrane signalling and activation...

Descripción completa

Detalles Bibliográficos
Autores principales: Alexander, J. Andrew N., Worrall, Liam J., Hu, Jinhong, Vuckovic, Marija, Satishkumar, Nidhi, Poon, Raymond, Sobhanifar, Solmaz, Rosell, Federico I., Jenkins, Joshua, Chiang, Daniel, Mosimann, Wesley A., Chambers, Henry F., Paetzel, Mark, Chatterjee, Som S., Strynadka, Natalie C. J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9834060/
https://www.ncbi.nlm.nih.gov/pubmed/36599987
http://dx.doi.org/10.1038/s41586-022-05583-3
_version_ 1784868376864096256
author Alexander, J. Andrew N.
Worrall, Liam J.
Hu, Jinhong
Vuckovic, Marija
Satishkumar, Nidhi
Poon, Raymond
Sobhanifar, Solmaz
Rosell, Federico I.
Jenkins, Joshua
Chiang, Daniel
Mosimann, Wesley A.
Chambers, Henry F.
Paetzel, Mark
Chatterjee, Som S.
Strynadka, Natalie C. J.
author_facet Alexander, J. Andrew N.
Worrall, Liam J.
Hu, Jinhong
Vuckovic, Marija
Satishkumar, Nidhi
Poon, Raymond
Sobhanifar, Solmaz
Rosell, Federico I.
Jenkins, Joshua
Chiang, Daniel
Mosimann, Wesley A.
Chambers, Henry F.
Paetzel, Mark
Chatterjee, Som S.
Strynadka, Natalie C. J.
author_sort Alexander, J. Andrew N.
collection PubMed
description Broad-spectrum β-lactam antibiotic resistance in Staphylococcus aureus is a global healthcare burden(1,2). In clinical strains, resistance is largely controlled by BlaR1(3), a receptor that senses β-lactams through the acylation of its sensor domain, inducing transmembrane signalling and activation of the cytoplasmic-facing metalloprotease domain(4). The metalloprotease domain has a role in BlaI derepression, inducing blaZ (β-lactamase PC1) and mecA (β-lactam-resistant cell-wall transpeptidase PBP2a) expression(3–7). Here, overcoming hurdles in isolation, we show that BlaR1 cleaves BlaI directly, as necessary for inactivation, with no requirement for additional components as suggested previously(8). Cryo-electron microscopy structures of BlaR1—the wild type and an autocleavage-deficient F284A mutant, with or without β-lactam—reveal a domain-swapped dimer that we suggest is critical to the stabilization of the signalling loops within. BlaR1 undergoes spontaneous autocleavage in cis between Ser283 and Phe284 and we describe the catalytic mechanism and specificity underlying the self and BlaI cleavage. The structures suggest that allosteric signalling emanates from β-lactam-induced exclusion of the prominent extracellular loop bound competitively in the sensor-domain active site, driving subsequent dynamic motions, including a shift in the sensor towards the membrane and accompanying changes in the zinc metalloprotease domain. We propose that this enhances the expulsion of autocleaved products from the active site, shifting the equilibrium to a state that is permissive of efficient BlaI cleavage. Collectively, this study provides a structure of a two-component signalling receptor that mediates action—in this case, antibiotic resistance—through the direct cleavage of a repressor.
format Online
Article
Text
id pubmed-9834060
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-98340602023-01-13 Structural basis of broad-spectrum β-lactam resistance in Staphylococcus aureus Alexander, J. Andrew N. Worrall, Liam J. Hu, Jinhong Vuckovic, Marija Satishkumar, Nidhi Poon, Raymond Sobhanifar, Solmaz Rosell, Federico I. Jenkins, Joshua Chiang, Daniel Mosimann, Wesley A. Chambers, Henry F. Paetzel, Mark Chatterjee, Som S. Strynadka, Natalie C. J. Nature Article Broad-spectrum β-lactam antibiotic resistance in Staphylococcus aureus is a global healthcare burden(1,2). In clinical strains, resistance is largely controlled by BlaR1(3), a receptor that senses β-lactams through the acylation of its sensor domain, inducing transmembrane signalling and activation of the cytoplasmic-facing metalloprotease domain(4). The metalloprotease domain has a role in BlaI derepression, inducing blaZ (β-lactamase PC1) and mecA (β-lactam-resistant cell-wall transpeptidase PBP2a) expression(3–7). Here, overcoming hurdles in isolation, we show that BlaR1 cleaves BlaI directly, as necessary for inactivation, with no requirement for additional components as suggested previously(8). Cryo-electron microscopy structures of BlaR1—the wild type and an autocleavage-deficient F284A mutant, with or without β-lactam—reveal a domain-swapped dimer that we suggest is critical to the stabilization of the signalling loops within. BlaR1 undergoes spontaneous autocleavage in cis between Ser283 and Phe284 and we describe the catalytic mechanism and specificity underlying the self and BlaI cleavage. The structures suggest that allosteric signalling emanates from β-lactam-induced exclusion of the prominent extracellular loop bound competitively in the sensor-domain active site, driving subsequent dynamic motions, including a shift in the sensor towards the membrane and accompanying changes in the zinc metalloprotease domain. We propose that this enhances the expulsion of autocleaved products from the active site, shifting the equilibrium to a state that is permissive of efficient BlaI cleavage. Collectively, this study provides a structure of a two-component signalling receptor that mediates action—in this case, antibiotic resistance—through the direct cleavage of a repressor. Nature Publishing Group UK 2023-01-04 2023 /pmc/articles/PMC9834060/ /pubmed/36599987 http://dx.doi.org/10.1038/s41586-022-05583-3 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Alexander, J. Andrew N.
Worrall, Liam J.
Hu, Jinhong
Vuckovic, Marija
Satishkumar, Nidhi
Poon, Raymond
Sobhanifar, Solmaz
Rosell, Federico I.
Jenkins, Joshua
Chiang, Daniel
Mosimann, Wesley A.
Chambers, Henry F.
Paetzel, Mark
Chatterjee, Som S.
Strynadka, Natalie C. J.
Structural basis of broad-spectrum β-lactam resistance in Staphylococcus aureus
title Structural basis of broad-spectrum β-lactam resistance in Staphylococcus aureus
title_full Structural basis of broad-spectrum β-lactam resistance in Staphylococcus aureus
title_fullStr Structural basis of broad-spectrum β-lactam resistance in Staphylococcus aureus
title_full_unstemmed Structural basis of broad-spectrum β-lactam resistance in Staphylococcus aureus
title_short Structural basis of broad-spectrum β-lactam resistance in Staphylococcus aureus
title_sort structural basis of broad-spectrum β-lactam resistance in staphylococcus aureus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9834060/
https://www.ncbi.nlm.nih.gov/pubmed/36599987
http://dx.doi.org/10.1038/s41586-022-05583-3
work_keys_str_mv AT alexanderjandrewn structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT worrallliamj structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT hujinhong structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT vuckovicmarija structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT satishkumarnidhi structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT poonraymond structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT sobhanifarsolmaz structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT rosellfedericoi structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT jenkinsjoshua structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT chiangdaniel structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT mosimannwesleya structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT chambershenryf structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT paetzelmark structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT chatterjeesoms structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus
AT strynadkanataliecj structuralbasisofbroadspectrumblactamresistanceinstaphylococcusaureus