High-Throughput Proteomics Identifies Proteins With Importance to Postantibiotic Recovery in Depolarized Persister Cells
Bacterial populations produce phenotypic variants called persisters to survive harmful conditions. Persisters are highly tolerant to antibiotics and repopulate environments after the stress has vanished. In order to resume growth, persisters have to recover from the persistent state, but the process...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-03-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.00378/full |
| _version_ | 1811270273087832064 |
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| author | Daniel-Timon Spanka Anne Konzer Daniel Edelmann Bork A. Berghoff |
| author_facet | Daniel-Timon Spanka Anne Konzer Daniel Edelmann Bork A. Berghoff |
| author_sort | Daniel-Timon Spanka |
| collection | DOAJ |
| description | Bacterial populations produce phenotypic variants called persisters to survive harmful conditions. Persisters are highly tolerant to antibiotics and repopulate environments after the stress has vanished. In order to resume growth, persisters have to recover from the persistent state, but the processes behind recovery remain mostly elusive. Deciphering these processes is an essential step toward understanding the persister phenomenon in its entirety. High-throughput proteomics by mass spectrometry is a valuable tool to assess persister physiology during any stage of the persister life cycle, and is expected to considerably contribute to our understanding of the recovery process. In the present study, an Escherichia coli strain, that overproduces the membrane-depolarizing toxin TisB, was established as a model for persistence by the use of high-throughput proteomics. Labeling of TisB persisters with stable isotope-containing amino acids (pulsed-SILAC) revealed an active translational response to ampicillin, including several RpoS-dependent proteins. Subsequent investigation of the persister proteome during postantibiotic recovery by label-free quantitative proteomics identified proteins with importance to the recovery process. Among them, AhpF, a component of alkyl hydroperoxide reductase, and the outer membrane porin OmpF were found to affect the persistence time of TisB persisters. Assessing the role of AhpF and OmpF in TisB-independent persisters demonstrated that the importance of a particular protein for the recovery process strongly depends on the physiological condition of a persister cell. Our study provides important insights into persister physiology and the processes behind recovery of depolarized cells. |
| first_indexed | 2024-04-12T21:57:49Z |
| format | Article |
| id | doaj.art-e801b653c4464f7e97e74fd5800722ea |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-12T21:57:49Z |
| publishDate | 2019-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-e801b653c4464f7e97e74fd5800722ea2022-12-22T03:15:15ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-03-011010.3389/fmicb.2019.00378424572High-Throughput Proteomics Identifies Proteins With Importance to Postantibiotic Recovery in Depolarized Persister CellsDaniel-Timon Spanka0Anne Konzer1Daniel Edelmann2Bork A. Berghoff3Institute for Microbiology and Molecular Biology, Justus Liebig University Giessen, Giessen, GermanyBiomolecular Mass Spectrometry, Max-Planck-Institute for Heart and Lung Research, Bad Nauheim, GermanyInstitute for Microbiology and Molecular Biology, Justus Liebig University Giessen, Giessen, GermanyInstitute for Microbiology and Molecular Biology, Justus Liebig University Giessen, Giessen, GermanyBacterial populations produce phenotypic variants called persisters to survive harmful conditions. Persisters are highly tolerant to antibiotics and repopulate environments after the stress has vanished. In order to resume growth, persisters have to recover from the persistent state, but the processes behind recovery remain mostly elusive. Deciphering these processes is an essential step toward understanding the persister phenomenon in its entirety. High-throughput proteomics by mass spectrometry is a valuable tool to assess persister physiology during any stage of the persister life cycle, and is expected to considerably contribute to our understanding of the recovery process. In the present study, an Escherichia coli strain, that overproduces the membrane-depolarizing toxin TisB, was established as a model for persistence by the use of high-throughput proteomics. Labeling of TisB persisters with stable isotope-containing amino acids (pulsed-SILAC) revealed an active translational response to ampicillin, including several RpoS-dependent proteins. Subsequent investigation of the persister proteome during postantibiotic recovery by label-free quantitative proteomics identified proteins with importance to the recovery process. Among them, AhpF, a component of alkyl hydroperoxide reductase, and the outer membrane porin OmpF were found to affect the persistence time of TisB persisters. Assessing the role of AhpF and OmpF in TisB-independent persisters demonstrated that the importance of a particular protein for the recovery process strongly depends on the physiological condition of a persister cell. Our study provides important insights into persister physiology and the processes behind recovery of depolarized cells.https://www.frontiersin.org/article/10.3389/fmicb.2019.00378/fullpersister cellsantibiotic tolerancedepolarizationTisB toxinrecoverySILAC |
| spellingShingle | Daniel-Timon Spanka Anne Konzer Daniel Edelmann Bork A. Berghoff High-Throughput Proteomics Identifies Proteins With Importance to Postantibiotic Recovery in Depolarized Persister Cells Frontiers in Microbiology persister cells antibiotic tolerance depolarization TisB toxin recovery SILAC |
| title | High-Throughput Proteomics Identifies Proteins With Importance to Postantibiotic Recovery in Depolarized Persister Cells |
| title_full | High-Throughput Proteomics Identifies Proteins With Importance to Postantibiotic Recovery in Depolarized Persister Cells |
| title_fullStr | High-Throughput Proteomics Identifies Proteins With Importance to Postantibiotic Recovery in Depolarized Persister Cells |
| title_full_unstemmed | High-Throughput Proteomics Identifies Proteins With Importance to Postantibiotic Recovery in Depolarized Persister Cells |
| title_short | High-Throughput Proteomics Identifies Proteins With Importance to Postantibiotic Recovery in Depolarized Persister Cells |
| title_sort | high throughput proteomics identifies proteins with importance to postantibiotic recovery in depolarized persister cells |
| topic | persister cells antibiotic tolerance depolarization TisB toxin recovery SILAC |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2019.00378/full |
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