Temperature-dependent regulation of bacterial cell division hydrolases by the coordinated action of a regulatory RNA and the ClpXP protease

A defining feature of bacteria is the peptidoglycan cell wall which provides structural integrity and prevents osmotic lysis. While peptidoglycan hydrolases are required for daughter cell separation, dysregulated cell wall degradation may result in cell lysis. The mechanisms allowing bacteria to con...

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Main Authors: Viktor H. Mebus, Larissa M. Busch, Morten Børre, Tobias K. Nielsen, Martin Saxtorph Bojer, Camilla Henriksen, Maria D. Barbuti, Danae M. Angeles, Kamilla Brejndal, Stephan Michalik, Manuela Gesell Salazar, Morten Kjos, Uwe Völker, Birgitte H. Kallipolitis, Dorte Frees
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:The Cell Surface
Online Access:http://www.sciencedirect.com/science/article/pii/S2468233025000040
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author Viktor H. Mebus
Larissa M. Busch
Morten Børre
Tobias K. Nielsen
Martin Saxtorph Bojer
Camilla Henriksen
Maria D. Barbuti
Danae M. Angeles
Kamilla Brejndal
Stephan Michalik
Manuela Gesell Salazar
Morten Kjos
Uwe Völker
Birgitte H. Kallipolitis
Dorte Frees
author_facet Viktor H. Mebus
Larissa M. Busch
Morten Børre
Tobias K. Nielsen
Martin Saxtorph Bojer
Camilla Henriksen
Maria D. Barbuti
Danae M. Angeles
Kamilla Brejndal
Stephan Michalik
Manuela Gesell Salazar
Morten Kjos
Uwe Völker
Birgitte H. Kallipolitis
Dorte Frees
author_sort Viktor H. Mebus
collection DOAJ
description A defining feature of bacteria is the peptidoglycan cell wall which provides structural integrity and prevents osmotic lysis. While peptidoglycan hydrolases are required for daughter cell separation, dysregulated cell wall degradation may result in cell lysis. The mechanisms allowing bacteria to control these deadly enzymes in response to environmental changes remain incompletely understood. Here, we find that in Staphylococcus aureus, temperature-dependent regulation of such hydrolases occurs by the coordinated action of a CHAP domain-specific regulatory RNA and the ClpXP protease. Using a proteomics approach, we identify a hitherto uncharacterized ClpXP controlled autolysin, CxcA, with a catalytic CHAP domain and show that it contributes to separation of daughter cells. CxcA is positively controlled by a non-coding RNA, named Rbc1 (for RNA binding to CHAP domain) transcribed from the antisense strand of cxcA. Notably, Rbc1 is capable of base pairing with RNAs encoding the CHAP domains of numerous cell wall hydrolases and we show that Rbc1 works in trans to upregulate the cell division hydrolase Sle1. Specifically, Rbc1 functions as a thermosensor allowing for upregulation of CxcA and Sle1 at low temperature where daughter cell separation is impeded. Interestingly, the Rbc1-mediated up-regulation of CxcA and Sle1 does not involve mRNA stabilization or increased translation; instead, Rbc1 depletion increases ClpXP-mediated degradation. In conclusion, we identify a novel cell division hydrolase that is highly conserved in Staphylococci and show that it is co-regulated with enzymes containing the catalytic CHAP domain via transcriptional regulation, an RNA-RNA temperature sensory mechanism and the ClpXP protease.
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spelling doaj-art-a5811dca49d14f3ea9946f85e49c03b52025-08-20T03:48:19ZengElsevierThe Cell Surface2468-23302025-06-011310014410.1016/j.tcsw.2025.100144Temperature-dependent regulation of bacterial cell division hydrolases by the coordinated action of a regulatory RNA and the ClpXP proteaseViktor H. Mebus0Larissa M. Busch1Morten Børre2Tobias K. Nielsen3Martin Saxtorph Bojer4Camilla Henriksen5Maria D. Barbuti6Danae M. Angeles7Kamilla Brejndal8Stephan Michalik9Manuela Gesell Salazar10Morten Kjos11Uwe Völker12Birgitte H. Kallipolitis13Dorte Frees14Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; Department of Biochemistry and Molecular Biology, Faculty of Natural Sciences, University of Southern Denmark, Odense, Denmark.Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Greifswald, GermanyDepartment of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, DenmarkDepartment of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, DenmarkDepartment of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, DenmarkDepartment of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, DenmarkFaculty of Chemistry, Biotechnology and Food Sciences, Norwegian University of Life Sciences, NorwayFaculty of Chemistry, Biotechnology and Food Sciences, Norwegian University of Life Sciences, NorwayDepartment of Biochemistry and Molecular Biology, Faculty of Natural Sciences, University of Southern Denmark, Odense, Denmark.Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Greifswald, GermanyDepartment of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Greifswald, GermanyFaculty of Chemistry, Biotechnology and Food Sciences, Norwegian University of Life Sciences, NorwayDepartment of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Greifswald, GermanyDepartment of Biochemistry and Molecular Biology, Faculty of Natural Sciences, University of Southern Denmark, Odense, Denmark.Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; Corresponding author at: Department of Veterinary and Animal Sciences, University of Copenhagen, Stigbøjlen 4, DK-1870 Frederiksberg C, Denmark.A defining feature of bacteria is the peptidoglycan cell wall which provides structural integrity and prevents osmotic lysis. While peptidoglycan hydrolases are required for daughter cell separation, dysregulated cell wall degradation may result in cell lysis. The mechanisms allowing bacteria to control these deadly enzymes in response to environmental changes remain incompletely understood. Here, we find that in Staphylococcus aureus, temperature-dependent regulation of such hydrolases occurs by the coordinated action of a CHAP domain-specific regulatory RNA and the ClpXP protease. Using a proteomics approach, we identify a hitherto uncharacterized ClpXP controlled autolysin, CxcA, with a catalytic CHAP domain and show that it contributes to separation of daughter cells. CxcA is positively controlled by a non-coding RNA, named Rbc1 (for RNA binding to CHAP domain) transcribed from the antisense strand of cxcA. Notably, Rbc1 is capable of base pairing with RNAs encoding the CHAP domains of numerous cell wall hydrolases and we show that Rbc1 works in trans to upregulate the cell division hydrolase Sle1. Specifically, Rbc1 functions as a thermosensor allowing for upregulation of CxcA and Sle1 at low temperature where daughter cell separation is impeded. Interestingly, the Rbc1-mediated up-regulation of CxcA and Sle1 does not involve mRNA stabilization or increased translation; instead, Rbc1 depletion increases ClpXP-mediated degradation. In conclusion, we identify a novel cell division hydrolase that is highly conserved in Staphylococci and show that it is co-regulated with enzymes containing the catalytic CHAP domain via transcriptional regulation, an RNA-RNA temperature sensory mechanism and the ClpXP protease.http://www.sciencedirect.com/science/article/pii/S2468233025000040
spellingShingle Viktor H. Mebus
Larissa M. Busch
Morten Børre
Tobias K. Nielsen
Martin Saxtorph Bojer
Camilla Henriksen
Maria D. Barbuti
Danae M. Angeles
Kamilla Brejndal
Stephan Michalik
Manuela Gesell Salazar
Morten Kjos
Uwe Völker
Birgitte H. Kallipolitis
Dorte Frees
Temperature-dependent regulation of bacterial cell division hydrolases by the coordinated action of a regulatory RNA and the ClpXP protease
The Cell Surface
title Temperature-dependent regulation of bacterial cell division hydrolases by the coordinated action of a regulatory RNA and the ClpXP protease
title_full Temperature-dependent regulation of bacterial cell division hydrolases by the coordinated action of a regulatory RNA and the ClpXP protease
title_fullStr Temperature-dependent regulation of bacterial cell division hydrolases by the coordinated action of a regulatory RNA and the ClpXP protease
title_full_unstemmed Temperature-dependent regulation of bacterial cell division hydrolases by the coordinated action of a regulatory RNA and the ClpXP protease
title_short Temperature-dependent regulation of bacterial cell division hydrolases by the coordinated action of a regulatory RNA and the ClpXP protease
title_sort temperature dependent regulation of bacterial cell division hydrolases by the coordinated action of a regulatory rna and the clpxp protease
url http://www.sciencedirect.com/science/article/pii/S2468233025000040
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