1887

Abstract

This study describes the identification and partial characterization of persistence-inducing factors (PIFs) from staphylococci.

Increases in persisters during mid-log phase growth indicate that quorum-sensing factors might be produced by staphylococci.

To identify and partially characterize PIFs from RP62A and SH1000.

Others have demonstrated a significant increase in persister numbers during mid-log phase. Inducers of this mid-log increase have yet to be identified in staphylococci. Optical density at 600 nm (OD) was used instead of time to determine when persister numbers increased during logarithmic growth. Concentrated culture filtrates (CCFs) from and were obtained at various ODs and following incubation at 16 h. The CCFs were used to develop a PIF assay. The PIF assay was used to partially characterize PIF from and for sizing of PIF activity, temperature and protease sensitivity and inter-species communications.

The optimal ODs for and PIF assays were 2.0 and 0.5, respectively. The highest PIF activity for both species was from CCF following incubation overnight (16 h). ’ PIF activity was decreased by storage at 4 C but not at 20 C (16 h), 37 C (1 h) or 100 C (15 min). ’ PIF activity was decreased following storage at 4 C (2 weeks) and after boiling at 100 C for 5 min but not after incubation at 37 C (1 h). PIF activity from both species went through a 3000 molecular weight cutoff ultrafilter. Proteinase K treatment of PIF decreased activity but did not decrease the PIF activity of . PIF from did not increase persisters when used to treat cells and nor did PIF from increase persisters when used to treat cells.

Attempts to discover PIFs for staphylococci were unsuccessful due to the time-based means used to identify mid-log. Both staphylococcal species produce extracellular, low-molecular-weight inducers of persistence when assayed using an OD -based PIF assay.

Funding
This study was supported by the:
  • A.T. Still University, Biomedical Sciences Master's Program
    • Principle Award Recipient: NealR Chamberlain
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2021-06-25
2024-04-26
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References

  1. Carroll KC, Butel JS, Mietzner TA. Chapter 13: The Staphylococci. In Jawetz, Melnick, and Adelberg’s Medical Microbiology, 27e, 27th. edn McGraw Hill; 2015
    [Google Scholar]
  2. Plata K, Rosato AE, Wegrzyn G. Staphylococcus aureus as an infectious agent: Overview of biochemistry and molecular genetics of its pathogenicity. Acta Biochim Pol 2009; 56:597–612 [View Article] [PubMed]
    [Google Scholar]
  3. Christensen GD, Simpson WA, Younger JJ et al. Adherence of coagulase-negative Staphylococci to plastic tissue culture plates: A quantitative model for the adherence of Staphylococci to medical devices. J Clin Microbiol 1985; 22:996–1006 [View Article] [PubMed]
    [Google Scholar]
  4. Shapiro JA, Nguyen VL, Chamberlain NR. Evidence for persisters in Staphylococcus epidermidis RP62a planktonic cultures and biofilms. J Med Microbiol 2011; 60:950–960 [View Article] [PubMed]
    [Google Scholar]
  5. Lewis K. eds Persister Cells and Infectious Disease Cham: Springer International Publishing; 2019 [View Article]
    [Google Scholar]
  6. Keren I, Kaldalu N, Spoering A, Wang Y, Lewis K. Persister cells and tolerance to antimicrobials. FEMS Microbiol Lett 2004; 230:13–18 [View Article] [PubMed]
    [Google Scholar]
  7. Hobby GL, Meyer K, Chaffee E. Observations on the mechanism of action of penicillin. Proc Soc Exp Biol Med 1942; 50:281–285 [View Article]
    [Google Scholar]
  8. Bigger J. Treatment of Staphylococcal infections with penicillin by intermittent sterilisation. Lancet 1944; 244:497–500 [View Article]
    [Google Scholar]
  9. Balaban NQ, Merrin J, Chait R, Kowalik L, Leibler S. Bacterial Persistance as a Phenotypic Switch 2004. Sci 2010; 10:305
    [Google Scholar]
  10. Balaban NQ, Merrin J, Chait R, Kowalik L, Leibler S. Bacterial persistence as a phenotypic switch. Sci 2004 [View Article]
    [Google Scholar]
  11. Lewis K. Persister cells, dormancy and infectious disease. Nat Rev Microbiol 2007; 5:48–56 [View Article] [PubMed]
    [Google Scholar]
  12. Fraikin N, Goormaghtigh F, Van Melderen L. Toxin-Antitoxin Systems and Persistence. In Persister Cells and Infectious Disease 2019 [View Article]
    [Google Scholar]
  13. Conlon BP, Rowe SE, Gandt AB et al. Persister formation in Staphylococcus aureus is associated with ATP depletion. Nat Microbiol 2016; 1: [View Article] [PubMed]
    [Google Scholar]
  14. Shan Y, Brown Gandt A, Rowe SE, Deisinger JP, Conlon BP et al. ATP-Dependent persister formation in Escherichia coli. mBio 2017; 8: [View Article]
    [Google Scholar]
  15. Fung DKC, Chan EWC, Chin ML, Chan RCY. Delineation of a bacterial starvation stress response network which can mediate antibiotic tolerance development. Antimicrob Agents Chemother 2010; 54:1082–1093 [View Article] [PubMed]
    [Google Scholar]
  16. Maisonneuve E, Shakespeare LJ, Jørgensen MG, Gerdes K. Bacterial persistence by RNA endonucleases. Proc Natl Acad Sci U S A 2011; 108:13206–13211 [View Article] [PubMed]
    [Google Scholar]
  17. Nguyen D, Joshi-Datar A, Lepine F et al. Active starvation responses mediate antibiotic tolerance in biofilms and nutrient-limited bacteria. Science 334:982–986 [View Article] [PubMed]
    [Google Scholar]
  18. Mordukhova EA, Pan JG. Stabilization of Homoserine-o-succinyltransferase (MetA) decreases the frequency of persisters in Escherichia coli under stressful conditions. PLoS One 2014; 9:e110504 [View Article] [PubMed]
    [Google Scholar]
  19. Murakami K, Ono T, Viducic D et al. Role for rpoS gene of Pseudomonas aeruginosa in antibiotic tolerance. FEMS Microbiol Lett 2005; 242:161–167 [View Article] [PubMed]
    [Google Scholar]
  20. Hong SH, Wang X, O’Connor HF, Benedik MJ, Wood TK. Bacterial persistence increases as environmental fitness decreases. Microb Biotechnol 2012; 5:509–522 [View Article] [PubMed]
    [Google Scholar]
  21. Wu Y, Vulić M, Keren I, Lewis K. Role of oxidative stress in persister tolerance. Antimicrob Agents Chemother 2012; 56:4922–4926 [View Article] [PubMed]
    [Google Scholar]
  22. Dörr T, Lewis K, Vulic M. SOS response induces persistence to fluoroquinolones in Escherichia coli. SEI Tech Rev 2007
    [Google Scholar]
  23. Johnson PJT, Levin BR. Pharmacodynamics, population dynamics, and the evolution of persistence in Staphylococcus aureus. PLoS Genet 2013; 9:e1003123 [View Article] [PubMed]
    [Google Scholar]
  24. Kwan BW, Valenta JA, Benedik MJ, Wood TK. Arrested protein synthesis increases persister-like cell formation. Antimicrob Agents Chemother 2013; 57:1468–1473 [View Article] [PubMed]
    [Google Scholar]
  25. Lewis K. Persister cells. Annu Rev Microbiol 2010; 64:357–372 [View Article] [PubMed]
    [Google Scholar]
  26. Möker N, Dean CR, Tao J. Pseudomonas aeruginosa increases formation of multidrug-tolerant persister cells in response to quorum-sensing signaling molecules. J Bacteriol 2010; 192:1946–1955 [View Article] [PubMed]
    [Google Scholar]
  27. Bhargava N, Sharma P, Capalash N. Pyocyanin stimulates quorum sensing-mediated tolerance to oxidative stress and increases persister cell populations in Acinetobacter baumannii. Infect Immun 2014; 82:3417–3425 [View Article] [PubMed]
    [Google Scholar]
  28. Leung V, Ajdic D, Koyanagi S, Lévesque CM. The formation of Streptococcus mutans persisters induced by the quorum-sensing peptide pheromone is affected by the LexA regulator. J Bacteriol 2015; 197:1083–1094 [View Article] [PubMed]
    [Google Scholar]
  29. Leung V, Lévesque CM. A stress-inducible quorum-sensing peptide mediates the formation of persister cells with noninherited multidrug tolerance. J Bacteriol 2012; 194:2265–2274 [View Article] [PubMed]
    [Google Scholar]
  30. Que YA, Hazan R, Strobel B et al. A quorum sensing small volatile molecule promotes antibiotic tolerance in bacteria. PLoS One 2013; 8:e80140 [View Article] [PubMed]
    [Google Scholar]
  31. Springer MT, Singh VK, Cheung AL, Donegan NP, Chamberlain NR. Effect of clpP and clpC deletion on persister cell number in Staphylococcus aureus. J Med Microbiol 2016; 65:848–857 [View Article] [PubMed]
    [Google Scholar]
  32. Baynes BM, Wang DIC, Trout BL. Role of arginine in the stabilization of proteins against aggregation. Biochemistry 2005; 44:4919–4925 [View Article] [PubMed]
    [Google Scholar]
  33. Ebeling W, Hennrich N, Klockow M, Metz H, Orth HD et al. Proteinase K from Tritirachium album Limber. Eur J Biochem 1974; 47:91–97 [View Article] [PubMed]
    [Google Scholar]
  34. Moyed HS, Broderick SH. Molecular cloning and expression of hipA, a gene of Escherichia coli K-12 that affects frequency of persistence after inhibition of murein synthesis. J Bacteriol 1986; 166:399–403 [View Article] [PubMed]
    [Google Scholar]
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