Publication

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

Wilmaerts, D., Bayoumi, M., Dewachter, L., Knapen, W., Mika, J. T., Hofkens, J., Dedecker, P., Maglia, G., Verstraeten, N. & Michiels, J., 2018, In : Mbio. 9, 4, 12 p., ARTN e00744-18.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Wilmaerts, D., Bayoumi, M., Dewachter, L., Knapen, W., Mika, J. T., Hofkens, J., ... Michiels, J. (2018). The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage. Mbio, 9(4), [ARTN e00744-18]. https://doi.org/10.1128/mBio.00744-18

Author

Wilmaerts, Dorien ; Bayoumi, Mariam ; Dewachter, Liselot ; Knapen, Wouter ; Mika, Jacek T ; Hofkens, Johan ; Dedecker, Peter ; Maglia, Giovanni ; Verstraeten, Natalie ; Michiels, Jan. / The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage. In: Mbio. 2018 ; Vol. 9, No. 4.

Harvard

Wilmaerts, D, Bayoumi, M, Dewachter, L, Knapen, W, Mika, JT, Hofkens, J, Dedecker, P, Maglia, G, Verstraeten, N & Michiels, J 2018, 'The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage', Mbio, vol. 9, no. 4, ARTN e00744-18. https://doi.org/10.1128/mBio.00744-18

Standard

The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage. / Wilmaerts, Dorien; Bayoumi, Mariam; Dewachter, Liselot; Knapen, Wouter; Mika, Jacek T; Hofkens, Johan; Dedecker, Peter; Maglia, Giovanni; Verstraeten, Natalie; Michiels, Jan.

In: Mbio, Vol. 9, No. 4, ARTN e00744-18, 2018.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Wilmaerts D, Bayoumi M, Dewachter L, Knapen W, Mika JT, Hofkens J et al. The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage. Mbio. 2018;9(4). ARTN e00744-18. https://doi.org/10.1128/mBio.00744-18


BibTeX

@article{4dbaec6f20c7487982f22d5df094a1a6,
title = "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 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 coil 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. coil 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 HokBinduced persister cells was strongly reduced using a channel blocker. The latter opens opportunities to reduce the number of persister cells in a clinical setting.",
keywords = "persistence, pore-forming peptide, toxin-antitoxin modules, PLANAR LIPID-BILAYERS, ESCHERICHIA-COLI, ANTITOXIN SYSTEMS, BACTERIAL PERSISTENCE, ANTIBIOTIC TOLERANCE, FLOW-CYTOMETRY, CELLS, EXPRESSION, MECHANISM, PROTEINS",
author = "Dorien Wilmaerts and Mariam Bayoumi and Liselot Dewachter and Wouter Knapen and Mika, {Jacek T} and Johan Hofkens and Peter Dedecker and Giovanni Maglia and Natalie Verstraeten and Jan Michiels",
note = "Copyright {\circledC} 2018 Wilmaerts et al.",
year = "2018",
doi = "10.1128/mBio.00744-18",
language = "English",
volume = "9",
journal = "Mbio",
issn = "2150-7511",
publisher = "AMER SOC MICROBIOLOGY",
number = "4",

}

RIS

TY - JOUR

T1 - The Persistence-Inducing Toxin HokB Forms Dynamic Pores That Cause ATP Leakage

AU - Wilmaerts, Dorien

AU - Bayoumi, Mariam

AU - Dewachter, Liselot

AU - Knapen, Wouter

AU - Mika, Jacek T

AU - Hofkens, Johan

AU - Dedecker, Peter

AU - Maglia, Giovanni

AU - Verstraeten, Natalie

AU - Michiels, Jan

N1 - Copyright © 2018 Wilmaerts et al.

PY - 2018

Y1 - 2018

N2 - 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 coil 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. coil 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 HokBinduced persister cells was strongly reduced using a channel blocker. The latter opens opportunities to reduce the number of persister cells in a clinical setting.

AB - 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 coil 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. coil 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 HokBinduced persister cells was strongly reduced using a channel blocker. The latter opens opportunities to reduce the number of persister cells in a clinical setting.

KW - persistence

KW - pore-forming peptide

KW - toxin-antitoxin modules

KW - PLANAR LIPID-BILAYERS

KW - ESCHERICHIA-COLI

KW - ANTITOXIN SYSTEMS

KW - BACTERIAL PERSISTENCE

KW - ANTIBIOTIC TOLERANCE

KW - FLOW-CYTOMETRY

KW - CELLS

KW - EXPRESSION

KW - MECHANISM

KW - PROTEINS

U2 - 10.1128/mBio.00744-18

DO - 10.1128/mBio.00744-18

M3 - Article

C2 - 30108166

VL - 9

JO - Mbio

JF - Mbio

SN - 2150-7511

IS - 4

M1 - ARTN e00744-18

ER -

ID: 64275097