Birth and Resuscitation of (p)ppGpp Induced Antibiotic Tolerant Persister Cells

Overview

Journal Sci Rep

Specialty Science

Date 2019 Apr 17

PMID 30988388

Citations 38

Authors

Mikkel Skjoldan Svenningsen

Alexandra Veress

Alexander Harms

Namiko Mitarai

Szabolcs Semsey

Affiliations

Soon will be listed here.

Abstract

Transient antibiotic treatment typically eradicates most sensitive bacteria except a few survivors called persisters. The second messenger (p)ppGpp plays a key role in persister formation in Escherichia coli populations but the underlying mechanisms have remained elusive. In this study we induced (p)ppGpp synthesis by modulating tRNA charging and then directly observed the stochastic appearance, antibiotic tolerance, and resuscitation of persister cells using live microscopy. Different physiological parameters of persister cells as well as their regularly growing ancestors and sisters were continuously monitored using fluorescent reporters. Our results confirmed previous findings that high (p)ppGpp levels are critical for persister formation, but the phenomenon remained strikingly stochastic without any correlation between (p)ppGpp levels and antibiotic tolerance on the single-cell level. We could not confirm previous notions that persisters exhibit markedly low concentrations of intracellular ATP or were linked to post-transcriptional effects of (p)ppGpp through the activation of small genetic elements known as toxin-antitoxin (TA) modules. Instead, we suggest that persister cell formation under regular conditions is driven by the transcriptional response to increased (p)ppGpp levels.

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References

Conlon B, Rowe S, Brown Gandt A, Nuxoll A, Donegan N, Zalis E . Persister formation in Staphylococcus aureus is associated with ATP depletion. Nat Microbiol. 2016; 1:16051. DOI: 10.1038/nmicrobiol.2016.51. View

Maisonneuve E, Castro-Camargo M, Gerdes K . (p)ppGpp Controls Bacterial Persistence by Stochastic Induction of Toxin-Antitoxin Activity. Cell. 2018; 172(5):1135. DOI: 10.1016/j.cell.2018.02.023. View

Yaginuma H, Kawai S, Tabata K, Tomiyama K, Kakizuka A, Komatsuzaki T . Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging. Sci Rep. 2014; 4:6522. PMC: 4185378. DOI: 10.1038/srep06522. View

Molina-Quiroz R, Silva-Valenzuela C, Brewster J, Castro-Nallar E, Levy S, Camilli A . Cyclic AMP Regulates Bacterial Persistence through Repression of the Oxidative Stress Response and SOS-Dependent DNA Repair in Uropathogenic . mBio. 2018; 9(1). PMC: 5760743. DOI: 10.1128/mBio.02144-17. View

Nguyen D, Joshi-Datar A, Lepine F, Bauerle E, Olakanmi O, Beer K . Active starvation responses mediate antibiotic tolerance in biofilms and nutrient-limited bacteria. Science. 2011; 334(6058):982-6. PMC: 4046891. DOI: 10.1126/science.1211037. View

Cataudella I, Sneppen K, Gerdes K, Mitarai N . Conditional cooperativity of toxin - antitoxin regulation can mediate bistability between growth and dormancy. PLoS Comput Biol. 2013; 9(8):e1003174. PMC: 3757081. DOI: 10.1371/journal.pcbi.1003174. View

Germain E, Roghanian M, Gerdes K, Maisonneuve E . Stochastic induction of persister cells by HipA through (p)ppGpp-mediated activation of mRNA endonucleases. Proc Natl Acad Sci U S A. 2015; 112(16):5171-6. PMC: 4413331. DOI: 10.1073/pnas.1423536112. View

Fitzpatrick A, Llabres S, Neuberger A, Blaza J, Bai X, Okada U . Structure of the MacAB-TolC ABC-type tripartite multidrug efflux pump. Nat Microbiol. 2017; 2:17070. PMC: 5447821. DOI: 10.1038/nmicrobiol.2017.70. View

Navarro Llorens J, Tormo A, Martinez-Garcia E . Stationary phase in gram-negative bacteria. FEMS Microbiol Rev. 2010; 34(4):476-95. DOI: 10.1111/j.1574-6976.2010.00213.x. View

10.

Maisonneuve E, Gerdes K . Molecular mechanisms underlying bacterial persisters. Cell. 2014; 157(3):539-48. DOI: 10.1016/j.cell.2014.02.050. View

11.

Aldridge B, Fernandez-Suarez M, Heller D, Ambravaneswaran V, Irimia D, Toner M . Asymmetry and aging of mycobacterial cells lead to variable growth and antibiotic susceptibility. Science. 2011; 335(6064):100-4. PMC: 3397429. DOI: 10.1126/science.1216166. View

12.

Chowdhury N, Kwan B, Wood T . Persistence Increases in the Absence of the Alarmone Guanosine Tetraphosphate by Reducing Cell Growth. Sci Rep. 2016; 6:20519. PMC: 4738310. DOI: 10.1038/srep20519. View

13.

Maisonneuve E, Castro-Camargo M, Gerdes K . (p)ppGpp controls bacterial persistence by stochastic induction of toxin-antitoxin activity. Cell. 2013; 154(5):1140-1150. DOI: 10.1016/j.cell.2013.07.048. View

14.

Pu Y, Zhao Z, Li Y, Zou J, Ma Q, Zhao Y . Enhanced Efflux Activity Facilitates Drug Tolerance in Dormant Bacterial Cells. Mol Cell. 2016; 62(2):284-294. PMC: 4850422. DOI: 10.1016/j.molcel.2016.03.035. View

15.

Radzikowski J, Vedelaar S, Siegel D, Ortega A, Schmidt A, Heinemann M . Bacterial persistence is an active σS stress response to metabolic flux limitation. Mol Syst Biol. 2016; 12(9):882. PMC: 5043093. DOI: 10.15252/msb.20166998. View

16.

Holden D, Errington J . Type II Toxin-Antitoxin Systems and Persister Cells. mBio. 2018; 9(5). PMC: 6156201. DOI: 10.1128/mBio.01574-18. View

17.

Balaban N, Merrin J, Chait R, Kowalik L, Leibler S . Bacterial persistence as a phenotypic switch. Science. 2004; 305(5690):1622-5. DOI: 10.1126/science.1099390. View

18.

Rocha E, Matic I, Taddei F . Over-representation of repeats in stress response genes: a strategy to increase versatility under stressful conditions?. Nucleic Acids Res. 2002; 30(9):1886-94. PMC: 113848. DOI: 10.1093/nar/30.9.1886. View

19.

Harms A, Fino C, Sorensen M, Semsey S, Gerdes K . Prophages and Growth Dynamics Confound Experimental Results with Antibiotic-Tolerant Persister Cells. mBio. 2017; 8(6). PMC: 5727415. DOI: 10.1128/mBio.01964-17. View

20.

Hoyland-Kroghsbo N, Maerkedahl R, Svenningsen S . A quorum-sensing-induced bacteriophage defense mechanism. mBio. 2013; 4(1):e00362-12. PMC: 3624510. DOI: 10.1128/mBio.00362-12. View