The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage

ABSTRACT Bacterial populations harbor a small fraction of cells that display transient multidrug tolerance. These so-called persister cells are extremely difficult to eradicate and contribute to the recalcitrance of chronic infections. Several signaling pathways leading to persistence have been iden...

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Autores principales:	Dorien Wilmaerts, Mariam Bayoumi, Liselot Dewachter, Wouter Knapen, Jacek T. Mika, Johan Hofkens, Peter Dedecker, Giovanni Maglia, Natalie Verstraeten, Jan Michiels
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	persistence pore-forming peptide toxin-antitoxin modules Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/8fd706619fea403d93938909da2131e9
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spelling	oai:doaj.org-article:8fd706619fea403d93938909da2131e92021-11-15T16:00:15ZThe Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage10.1128/mBio.00744-182150-7511https://doaj.org/article/8fd706619fea403d93938909da2131e92018-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00744-18https://doaj.org/toc/2150-7511ABSTRACT Bacterial populations harbor a small fraction of cells that display transient multidrug tolerance. These so-called persister cells are extremely difficult to eradicate and contribute to the recalcitrance of chronic infections. Several signaling pathways leading to persistence have been identified. However, it is poorly understood how the effectors of these pathways function at the molecular level. In a previous study, we reported that the conserved GTPase Obg induces persistence in Escherichia coli via transcriptional upregulation of the toxin HokB. In the present study, we demonstrate that HokB inserts in the cytoplasmic membrane where it forms pores. The pore-forming capacity of the HokB peptide is demonstrated by in vitro conductance measurements on synthetic and natural lipid bilayers, revealing an asymmetrical conductance profile. Pore formation is directly linked to persistence and results in leakage of intracellular ATP. HokB-induced persistence is strongly impeded in the presence of a channel blocker, thereby providing a direct link between pore functioning and persistence. Furthermore, the activity of HokB pores is sensitive to the membrane potential. This sensitivity presumably results from the formation of either intermediate or mature pore types depending on the membrane potential. Taken together, these results provide a detailed view on the mechanistic basis of persister formation through the effector HokB. IMPORTANCE There is increasing awareness of the clinical importance of persistence. Indeed, persistence is linked to the recalcitrance of chronic infections, and evidence is accumulating that persister cells constitute a pool of viable cells from which resistant mutants can emerge. Unfortunately, persistence is a poorly understood process at the mechanistic level. In this study, we unraveled the pore-forming activity of HokB in E. coli and discovered that these pores lead to leakage of intracellular ATP, which is correlated with the induction of persistence. Moreover, we established a link between persistence and pore activity, as the number of HokB-induced persister cells was strongly reduced using a channel blocker. The latter opens opportunities to reduce the number of persister cells in a clinical setting.Dorien WilmaertsMariam BayoumiLiselot DewachterWouter KnapenJacek T. MikaJohan HofkensPeter DedeckerGiovanni MagliaNatalie VerstraetenJan MichielsAmerican Society for Microbiologyarticlepersistencepore-forming peptidetoxin-antitoxin modulesMicrobiologyQR1-502ENmBio, Vol 9, Iss 4 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	persistence pore-forming peptide toxin-antitoxin modules Microbiology QR1-502
spellingShingle	persistence pore-forming peptide toxin-antitoxin modules Microbiology QR1-502 Dorien Wilmaerts Mariam Bayoumi Liselot Dewachter Wouter Knapen Jacek T. Mika Johan Hofkens Peter Dedecker Giovanni Maglia Natalie Verstraeten Jan Michiels The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage
description	ABSTRACT Bacterial populations harbor a small fraction of cells that display transient multidrug tolerance. These so-called persister cells are extremely difficult to eradicate and contribute to the recalcitrance of chronic infections. Several signaling pathways leading to persistence have been identified. However, it is poorly understood how the effectors of these pathways function at the molecular level. In a previous study, we reported that the conserved GTPase Obg induces persistence in Escherichia coli via transcriptional upregulation of the toxin HokB. In the present study, we demonstrate that HokB inserts in the cytoplasmic membrane where it forms pores. The pore-forming capacity of the HokB peptide is demonstrated by in vitro conductance measurements on synthetic and natural lipid bilayers, revealing an asymmetrical conductance profile. Pore formation is directly linked to persistence and results in leakage of intracellular ATP. HokB-induced persistence is strongly impeded in the presence of a channel blocker, thereby providing a direct link between pore functioning and persistence. Furthermore, the activity of HokB pores is sensitive to the membrane potential. This sensitivity presumably results from the formation of either intermediate or mature pore types depending on the membrane potential. Taken together, these results provide a detailed view on the mechanistic basis of persister formation through the effector HokB. IMPORTANCE There is increasing awareness of the clinical importance of persistence. Indeed, persistence is linked to the recalcitrance of chronic infections, and evidence is accumulating that persister cells constitute a pool of viable cells from which resistant mutants can emerge. Unfortunately, persistence is a poorly understood process at the mechanistic level. In this study, we unraveled the pore-forming activity of HokB in E. coli and discovered that these pores lead to leakage of intracellular ATP, which is correlated with the induction of persistence. Moreover, we established a link between persistence and pore activity, as the number of HokB-induced persister cells was strongly reduced using a channel blocker. The latter opens opportunities to reduce the number of persister cells in a clinical setting.
format	article
author	Dorien Wilmaerts Mariam Bayoumi Liselot Dewachter Wouter Knapen Jacek T. Mika Johan Hofkens Peter Dedecker Giovanni Maglia Natalie Verstraeten Jan Michiels
author_facet	Dorien Wilmaerts Mariam Bayoumi Liselot Dewachter Wouter Knapen Jacek T. Mika Johan Hofkens Peter Dedecker Giovanni Maglia Natalie Verstraeten Jan Michiels
author_sort	Dorien Wilmaerts
title	The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage
title_short	The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage
title_full	The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage
title_fullStr	The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage
title_full_unstemmed	The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage
title_sort	persistence-inducing toxin hokb forms dynamic pores that cause atp leakage
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/8fd706619fea403d93938909da2131e9
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The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage

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