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DNA susceptibility of Saccharomyces cerevisiae to Zeocin depends on the growth phase

  • Teodora Todorova
  • Daniela Miteva
  • Stephka ChankovaEmail author
Original Article

Abstract

The aim of this study was to evaluate the level of Zeocin-induced double-strand breaks (DSBs) in Saccharomyces cerevisiae cells in a different growth phase, using constant-field gel electrophoresis (CFGE). Saccharomyces cerevisiae diploid strain D7ts1 with enhanced cellular permeability was used. The effects of growth phase and treatment time were evaluated based on Zeocin-induced DSBs, measured by CFGE. Survival assay was also applied. No protoplast isolation was necessary for the detection of DSBs in strain D7ts1. Differences in the response of cells depending on the growth phase were obtained. Cells in exponential growth phase had increased DSB levels only after Zeocin treatment with concentrations equal or higher than 200 μgml−1. Increasing treatment time did not result in higher DSB levels. Oppositely, treatment of cells at the beginning of stationary phase with Zeocin concentrations resulted in more than 1.5-fold increase in DSB levels in comparison with those in untreated cells. Increased DSB levels were measured for all the treatment times. A dose-dependent decrease in cell survival was observed after Zeocin treatment with concentrations in the range of lethality LD20–LD50. A strong negative correlation was calculated between the levels of DSBs and cell survival. New information is provided concerning DNA susceptibility depending on the growth phase. DNA susceptibility is higher in cells at the beginning of stationary phase than those in exponential phase. Data presented here illustrate that the optimized by us CFGE protocol is sensitive and could be used successfully for DSB measurement in Saccharomyces cerevisiae strains with enhanced cellular permeability.

Keywords

CFGE DNA susceptibility Growth phase Saccharomyces cerevisiae Zeocin 

Notes

Funding information

This work was supported by a grant from the National Science Fund, Ministry of Education and Science, Project No. DH11/10.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Beam CA, Mortimer RK, Wolfe RG, Tobias CA (1954) The relation of radioresistance to budding in Saccharomyces cerevisiae. Arch Biochem Biophys 49:110–122CrossRefGoogle Scholar
  2. Bryant PE (1968) Survival after fractionated doses of radiation: modification by anoxia of the response of Chlamydomonas. Nature 219:75–77.  https://doi.org/10.1038/219075b0 CrossRefGoogle Scholar
  3. Chankova SG, Bryant PE (2002) Acceleration of DNA-double strand rejoining during the adaptive response of Chlamydomonas reinhardtii. Radiats Biol Radioecol 42(6):600–603Google Scholar
  4. Chankova SG, Matos JA, Simoes F, Bryant PE (2005) The adaptive response of a new radio-resistant strain of Chlamydomonas reinhardtii correlates with increased DNA double-strand break rejoining. Int J Radiat Biol 81(7):509–514CrossRefGoogle Scholar
  5. Chankova SG, Dimova E, Dimitrova M, Bryant PE (2007) Induction of DNA double-strand breaks by Zeocin in Chlamydomonas reinhardtii and the role of increased DNA double-strand breaks rejoining in the formation of an adaptive response. Radiat Environ Biophys 46:409–416CrossRefGoogle Scholar
  6. Chankova S, Todorova T, Parvanova P, Miteva D, Mitrovska Z, Angelova O, Imreova P, Mucaji P (2013) Kaempferol and jatropham: are they protective or detrimental for Chlamydomonas reinhardtii? C R Acad Bulg Sci 66:1121–1128Google Scholar
  7. Dimitrov MD, Pesheva MG, Venkov PV (2013) New cell-based assay indicates dependence of antioxidant biological the origin of reactive oxygen species. J Agric Food Chem 61:4344–4351CrossRefGoogle Scholar
  8. Dimova E, Dimitrova M, Miteva D, Mitrovska Z, Bryant PE, Chankova S (2009) Does single-dose cell resistance to the radiomimetic Zeocin correlate with a Zeocin-induced adaptive response in Chlamydomonas reinhardtii strains? Radiat Environ Biophys 48:77–84CrossRefGoogle Scholar
  9. Frassinetti S, Barberio C, Caltavuturo L, Fava F, Di Gioia D (2011) Genotoxicity of 4-nonylphenol and nonylphenol ethoxylate mixtures by the use of Saccharomyces cerevisiae D7 mutation assay and use of this text to evaluate the efficiency of biodegradation treatments. Ecotoxicol Environ Saf 74(3):253–258CrossRefGoogle Scholar
  10. Freeman KM, Hoffmann GR (2007) Frequencies of mutagen-induced coincident mitotic recombination at unlinked loci in Saccharomyces cerevisiae. Mutat Res Fundam Mol Mech Mutagen 616(1):119–132CrossRefGoogle Scholar
  11. Galao RP, Scheller N, Alves-Rodrigues I, Breinig T, Meyerhans A, Díez J (2007) Saccharomyces cerevisiae: a versatile eukaryotic system in virology. Microb Cell Factories 6:32CrossRefGoogle Scholar
  12. Gateva S, Angelova O, Chankova S (2015) Double-strand breaks detection in human lymphocytes by constant field gel electrophoresis. ДОКЛАДИ НА БЪЛГАРСКАТА АКАДЕМИЯ НА НАУКИТЕ 68(4):469–474Google Scholar
  13. Gray JV, Petsko GA, Johnston GC, Ringe D, Singer RA, Werner-Washburne M (2004) Sleeping beauty: quiescence in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 68(2):187–206CrossRefGoogle Scholar
  14. Hafer K, Rivina L, Schiestl RH (2009) Cell cycle dependence of ionizing radiation-induced DNA deletions and antioxidant radioprotection in Saccharomyces cerevisiae. Radiat Res 173(6):802–808CrossRefGoogle Scholar
  15. Holroyd LF, van Mourik T (2015) Stacking of the mutagenic base analogue 5-bromouracil: energy landscapes of pyrimidine dimers in gas phase and water. Phys Chem Chem Phys 17(45):30364–30370CrossRefGoogle Scholar
  16. Ikner A, Shiozaki K (2005) Yeast signaling pathways in the oxidative stress response. Mutat Res Fundam Mol Mech Mutagen 569(1-2):13–27CrossRefGoogle Scholar
  17. Kepes F, Schekman R (1988) The yeast SEC53 gene encodes phosphomannomutase. J Biol Chem 263(19):9155–9161Google Scholar
  18. Kopaskova M, Hadjo L, Yankulova B, Jovtchev G, Galova E, Sevcovicova A, Mucaji P, Miadokova E, Bryant P, Chankova S (2011) Extract of Lillium candidum L. can modulate the genotoxicity of the antibiotic Zeocin. Molecules 17:80–97CrossRefGoogle Scholar
  19. Kumar R, Srivastava S (2016) Quantitative proteomic comparison of stationary/G0 phase cells and tetrads in budding yeast. Sci Rep 6:32031CrossRefGoogle Scholar
  20. Langguth EN, Beam CA (1973) Repair mechanisms and cell cycle dependent variations in X-ray sensitivity of diploid yeast. Radiat Res 53:226–234CrossRefGoogle Scholar
  21. Lee Y, Kim K, Kang KT, Lee JS, Yang SS, Chung WH (2014) Atmospheric-pressure plasma jet induces DNA double-strand breaks that require a Rad51-mediated homologous recombination for repair in Saccharomyces cerevisiae. Arch Biochem Biophys 560:1–9CrossRefGoogle Scholar
  22. Lewis K (2000) Programmed death in bacteria. Microbiol Mol Biol Rev 64:503–533CrossRefGoogle Scholar
  23. Lim ST, Jue CK, Moore CW, Lipke PN (1995) Oxidative cell wall damage mediated by bleomycin-Fe (II) in Saccharomyces cerevisiae. J Bacteriol 177(12):3534–3539CrossRefGoogle Scholar
  24. López-Larraza D, De Luca JC, Bianchi NO (1990) The kinetics of DNA damage by bleomycin in mammalian cells. Mutat Res Fundam Mol Mech Mutagen 232(1):57–61CrossRefGoogle Scholar
  25. Lundin C, North M, Erixon K, Walters K, Jenssen D, Goldman AS, Helleday T (2005) Methyl methanesulfonate (MMS) produces heat-labile DNA damage but no detectable in vivo DNA double-strand breaks. Nucleic Acids Res 33(12):3799–3811CrossRefGoogle Scholar
  26. Matuo R, Sousa FG, Soares DG, Bonatto D, Saffi J, Escargueil AE, Larsen AK, Henriques JAP (2012) Saccharomyces cerevisiae as a model system to study the response to anticancer agents. Cancer Chemother Pharmacol 70(4):491–502CrossRefGoogle Scholar
  27. Methyl methanesulfonate (2018) https://pubchem.ncbi.nlm.nih.gov/compound/methyl_methanesulfonate#section=Top. Accessed 19 Dec 2018
  28. Moore CW (1982) Modulation of in vivo bleomycin cytotoxicity. Antimicrob Agents Chemother 21:595–600CrossRefGoogle Scholar
  29. Moore CW, Malcolm AW, Tomkinson KN, Little JB (1985) Ultrarapid recovery from lethal effects of bleomycin and γ-radiation in stationary-phase human diploid fibroblasts. Cancer Res 45(5):1978–1981Google Scholar
  30. Moore CW, Jones CS, Wall LA (1989) Growth phase dependency of chromatin cleavage and degradation by bleomycin. Antimicrob Agents Chemother 33(9):1592–1599CrossRefGoogle Scholar
  31. Moore CW, McKoy J, Dardalhon M, Davermann D, Martinez M, Averbeck D (2000) DNA damage-inducible and RAD52-independent repair of DNA double-strand breaks in Saccharomyces cerevisiae. Genetics 154:1085–1099Google Scholar
  32. Pellacani C, Buschini A, Furlini M, Poli P, Rossi C (2006) A battery of in vivo and in vitro tests useful for genotoxic pollutant detection in surface waters. Aquat Toxicol 77(1):1–10CrossRefGoogle Scholar
  33. Pesheva M, Krastanova O, Staleva L, Dentcheva V, Hadzhitodorov M, Venkov P (2005) The Ty1 transposition assay: a new short-term test for detection of carcinogens. J Microbiol Methods 61(1):1–8CrossRefGoogle Scholar
  34. Petin VG, Kapultcevich YG (2014) Radiation quality and the shape of dose–effect curves at low doses of ionizing radiation for eukaryotic cells. Math Biosci 252:1–6CrossRefGoogle Scholar
  35. Poli P, Buschini A, Campanini N, Vettori MV, Cassoni F, Cattani S, Rossi C (1992) Urban air pollution: use of different mutagenicity assays to evaluate environmental genetic hazard. Mutat Res 298:113–123CrossRefGoogle Scholar
  36. Poli P, de Mello MA, Buschini A, Mortara RA, de Albuquerque CN, da Silva S, Rossi C, Zucchi TMAD (2002) Cytotoxic and genotoxic effects of megazol, an anti-Chagas’ disease drug, assessed by different short-term tests. Biochem Pharmacol 64(11):1617–1627CrossRefGoogle Scholar
  37. Raju MR, Gnanapurani M, Stackler B, Madhvanath U, Howard J, Lyman JT, Manney TR, Tobias CA (1972) Influence of linear energy transfer on the radioresistance of budding haploid yeast cells. Radiat Res 51:310–317CrossRefGoogle Scholar
  38. Rao KS (2007) Mechanisms of disease: DNA repair defects and neurological disease. Nat Clin Pract Neurol 3:162–172CrossRefGoogle Scholar
  39. Reed LJ, Muench H (1938) A simple method of estimating fifty per cent endpoints. Am J Epidemiol 27(3):493–497CrossRefGoogle Scholar
  40. Rossi C, Poli P, Buschini A, Cassoni F, Cattani S, de Munari E (1995) Comparative investigations among meteorological conditions, air chemical–physical pollutants and airborne particulate mutagenicity: a long-term study (1990–1994) from a northern Italian town. Chemosphere 30:1829–1845CrossRefGoogle Scholar
  41. Sousa-Lopes A, Antunes F, Cyrne L, Marinho HS (2004) Decreased cellular permeability to H2O2 protects Saccharomyces cerevisiae cells in stationary phase against oxidative stress. FEBS Lett 578(1):152–156CrossRefGoogle Scholar
  42. Staleva L, Waltscheva L, Golovinsky E, Venkov P (1996) Enhanced cell permeability increases the sensitivity of a yeast test for mutagens. Mutat Res 370:81–89CrossRefGoogle Scholar
  43. Stoycheva T, Pesheva M, Dimitrov M, Venkov P (2012) The Ty1 retrotransposition short-term test for selective detection of carcinogenic genotoxins. In: Pesheva M (Ed) Carcinogen, Intechopen, pp 83–110Google Scholar
  44. Temple MD, Perrone GG, Dawes IW (2005) Complex cellular responses to reactive oxygen species. Trends Cell Biol 15(6):319–326CrossRefGoogle Scholar
  45. Tippins RS, Parry JM (1982) A comparison of the radiosensitivity of stationary, exponential, and G1 phase wild type and repair deficient yeast cultures: supporting evidence for stationary phase yeast cells being in G0. Int J Radiat Biol 41:215–220Google Scholar
  46. Todorova T, Pesheva M, Gregan F, Chankova S (2015a) Antioxidant, antimutagenic and anticarcinogenic effects of Papaver rhoeas L. extract on Saccharomyces cerevisiae. J Med Food 18(4):460–467CrossRefGoogle Scholar
  47. Todorova T, Miteva D, Chankova S (2015b) DNA damaging effect of Zeocin and methyl methanesulfonate in Saccharomyces cerevisiae measured by CFGE. ДОКЛАДИ НА БЪЛГАРСКАТА АКАДЕМИЯ НА НАУКИТЕ 68(1):71–78 Google Scholar
  48. Venkov P, Scheit KH (1984) Effect of seminal plasmin on rRNA synthesis in Saccharomyces cerevisiae. FEBS Lett 172(1):21–24CrossRefGoogle Scholar
  49. Werner-Washburne M, Braun E, Johnston GC, Singer RA (1993) Stationary phase in the yeast Saccharomyces cerevisiae. Microbiol Rev 57:383–401Google Scholar
  50. Wilson MR (2014) Analysis of genes required for quiescent cell formation in stationary phase cultures of Saccharomyces cerevisiae. Dissertation, University of New MexicoGoogle Scholar
  51. Zaka R, Chenal C (2002) Misset MT study of external low irradiation dose effects on induction of chromosome aberrations in Pisum sativum root tip meristem. Mut Res 517:87–99CrossRefGoogle Scholar
  52. Zimmermann FK, Kern R, Rasenberger H (1975) A yeast strain for simultaneous detection of induced mitotic crossing over, mitotic gene conversion and reverse mutation. Mutat Res Fundam Mol Mech Mutagen 28(3):381–388CrossRefGoogle Scholar
  53. Zlotnik KH, Femande MP, Bowers B, Cabib E (1989) Mannoproteins form an external cell wall layer determines wall porosity in Saccharomyces cerevisiae. J Bacteriol 195:1018–1026Google Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Teodora Todorova
    • 1
  • Daniela Miteva
    • 1
  • Stephka Chankova
    • 1
    Email author
  1. 1.Institute of Biodiversity and Ecosystem ResearchBulgarian Academy of SciencesSofiaBulgaria

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