Evidence That VirS Is a Receptor for the Signaling Peptide of the <named-content content-type="genus-species">Clostridium perfringens</named-content> Agr-like Quorum Sensing System

ABSTRACT Since both the Agr (accessory gene regulator)-like quorum sensing (QS) system and VirS/VirR (VirS/R) two-component regulatory system of Clostridium perfringens positively regulate production of several toxins, including C. perfringens beta toxin (CPB), it has been hypothesized the VirS memb...

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Autores principales:	Jihong Li, Bruce A. McClane
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Clostridium perfringens beta toxin Agr-like quorum sensing VirS/R two-component regulatory system signal peptide receptor toxin production regulation Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/be737479f52e410fbb61966eaf677c5d
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spelling	oai:doaj.org-article:be737479f52e410fbb61966eaf677c5d2021-11-15T16:19:07ZEvidence That VirS Is a Receptor for the Signaling Peptide of the <named-content content-type="genus-species">Clostridium perfringens</named-content> Agr-like Quorum Sensing System10.1128/mBio.02219-202150-7511https://doaj.org/article/be737479f52e410fbb61966eaf677c5d2020-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02219-20https://doaj.org/toc/2150-7511ABSTRACT Since both the Agr (accessory gene regulator)-like quorum sensing (QS) system and VirS/VirR (VirS/R) two-component regulatory system of Clostridium perfringens positively regulate production of several toxins, including C. perfringens beta toxin (CPB), it has been hypothesized the VirS membrane sensor protein is an Agr-like QS signaling peptide (SP) receptor. To begin evaluating whether VirS is an SP receptor, this study sequenced the virS gene in C. perfringens strains CN3685 and CN1795 because it was reported that agrB mutants of both strains increase CPB production in response to the pentapeptide 5R, likely the natural SP, but only the CN3685 agrB mutant responds to 8R, which is 5R plus a 3-amino-acid tail. This sequencing identified differences between the predicted VirS extracellular loop 2 (ECL2) of CN3685 versus that of CN1795. To explore if those ECL2 differences explain strain-related variations in SP sensitivity and support VirS as an SP receptor, virS agrB double-null mutants of each strain were complemented to swap which VirS protein they produce. CPB Western blotting showed that this complementation changed the natural responsiveness of each strain to 8R. A pulldown experiment using biotin-5R demonstrated that VirS can bind SP. To further support VirS:SP binding and to identify a VirS binding site for SP, a 14-mer peptide corresponding to VirS ECL2 was synthesized. This ECL2 peptide inhibited 5R signaling to agrB mutant and wild-type strains. This inhibition was specific, since a single N to D substitution in the ECL2 peptide abrogated these effects. Collectively, these results support VirS as an important SP receptor and may assist development of therapeutics. IMPORTANCE C. perfringens beta toxin (CPB) is essential for the virulence of type C strains, a common cause of fatal necrotizing enteritis and enterotoxemia in humans and domestic animals. Production of CPB, as well as several other C. perfringens toxins, is positively regulated by both the Agr-like QS system and the VirS/R two-component regulatory system. This study presents evidence that the VirS membrane sensor protein is a receptor for the AgrD-derived SP and that the second extracellular loop of VirS is important for SP binding. Understanding interactions between SP and VirS improves knowledge of C. perfringens pathogenicity and may provide insights for designing novel strategies to reduce C. perfringens toxin production during infections.Jihong LiBruce A. McClaneAmerican Society for MicrobiologyarticleClostridium perfringensbeta toxinAgr-like quorum sensingVirS/R two-component regulatory systemsignal peptide receptortoxin production regulationMicrobiologyQR1-502ENmBio, Vol 11, Iss 5 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Clostridium perfringens beta toxin Agr-like quorum sensing VirS/R two-component regulatory system signal peptide receptor toxin production regulation Microbiology QR1-502
spellingShingle	Clostridium perfringens beta toxin Agr-like quorum sensing VirS/R two-component regulatory system signal peptide receptor toxin production regulation Microbiology QR1-502 Jihong Li Bruce A. McClane Evidence That VirS Is a Receptor for the Signaling Peptide of the <named-content content-type="genus-species">Clostridium perfringens</named-content> Agr-like Quorum Sensing System
description	ABSTRACT Since both the Agr (accessory gene regulator)-like quorum sensing (QS) system and VirS/VirR (VirS/R) two-component regulatory system of Clostridium perfringens positively regulate production of several toxins, including C. perfringens beta toxin (CPB), it has been hypothesized the VirS membrane sensor protein is an Agr-like QS signaling peptide (SP) receptor. To begin evaluating whether VirS is an SP receptor, this study sequenced the virS gene in C. perfringens strains CN3685 and CN1795 because it was reported that agrB mutants of both strains increase CPB production in response to the pentapeptide 5R, likely the natural SP, but only the CN3685 agrB mutant responds to 8R, which is 5R plus a 3-amino-acid tail. This sequencing identified differences between the predicted VirS extracellular loop 2 (ECL2) of CN3685 versus that of CN1795. To explore if those ECL2 differences explain strain-related variations in SP sensitivity and support VirS as an SP receptor, virS agrB double-null mutants of each strain were complemented to swap which VirS protein they produce. CPB Western blotting showed that this complementation changed the natural responsiveness of each strain to 8R. A pulldown experiment using biotin-5R demonstrated that VirS can bind SP. To further support VirS:SP binding and to identify a VirS binding site for SP, a 14-mer peptide corresponding to VirS ECL2 was synthesized. This ECL2 peptide inhibited 5R signaling to agrB mutant and wild-type strains. This inhibition was specific, since a single N to D substitution in the ECL2 peptide abrogated these effects. Collectively, these results support VirS as an important SP receptor and may assist development of therapeutics. IMPORTANCE C. perfringens beta toxin (CPB) is essential for the virulence of type C strains, a common cause of fatal necrotizing enteritis and enterotoxemia in humans and domestic animals. Production of CPB, as well as several other C. perfringens toxins, is positively regulated by both the Agr-like QS system and the VirS/R two-component regulatory system. This study presents evidence that the VirS membrane sensor protein is a receptor for the AgrD-derived SP and that the second extracellular loop of VirS is important for SP binding. Understanding interactions between SP and VirS improves knowledge of C. perfringens pathogenicity and may provide insights for designing novel strategies to reduce C. perfringens toxin production during infections.
format	article
author	Jihong Li Bruce A. McClane
author_facet	Jihong Li Bruce A. McClane
author_sort	Jihong Li
title	Evidence That VirS Is a Receptor for the Signaling Peptide of the <named-content content-type="genus-species">Clostridium perfringens</named-content> Agr-like Quorum Sensing System
title_short	Evidence That VirS Is a Receptor for the Signaling Peptide of the <named-content content-type="genus-species">Clostridium perfringens</named-content> Agr-like Quorum Sensing System
title_full	Evidence That VirS Is a Receptor for the Signaling Peptide of the <named-content content-type="genus-species">Clostridium perfringens</named-content> Agr-like Quorum Sensing System
title_fullStr	Evidence That VirS Is a Receptor for the Signaling Peptide of the <named-content content-type="genus-species">Clostridium perfringens</named-content> Agr-like Quorum Sensing System
title_full_unstemmed	Evidence That VirS Is a Receptor for the Signaling Peptide of the <named-content content-type="genus-species">Clostridium perfringens</named-content> Agr-like Quorum Sensing System
title_sort	evidence that virs is a receptor for the signaling peptide of the <named-content content-type="genus-species">clostridium perfringens</named-content> agr-like quorum sensing system
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/be737479f52e410fbb61966eaf677c5d
work_keys_str_mv	AT jihongli evidencethatvirsisareceptorforthesignalingpeptideofthenamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentagrlikequorumsensingsystem AT bruceamcclane evidencethatvirsisareceptorforthesignalingpeptideofthenamedcontentcontenttypegenusspeciesclostridiumperfringensnamedcontentagrlikequorumsensingsystem
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Evidence That VirS Is a Receptor for the Signaling Peptide of the <named-content content-type="genus-species">Clostridium perfringens</named-content> Agr-like Quorum Sensing System

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