Advertisement

Apoptosis

, Volume 21, Issue 7, pp 866–872 | Cite as

The fungicide Mancozeb induces metacaspase-dependent apoptotic cell death in Saccharomyces cerevisiae BY4741

  • F. J. Scariot
  • L. M. Jahn
  • J. P. Maianti
  • A. P. L. Delamare
  • S. EcheverrigarayEmail author
Article

Abstract

Mancozeb (MZ), a mixture of ethylene-bis-dithiocarbamate manganese and zinc salts, is one of the most widely used fungicides in agriculture. Toxicologic studies in mammals and mammalian cells indicate that this fungicide can cause neurological and cytological disorders, putatively associated with pro-oxidant and apoptotic effects. Yeast adaptation to sub-inhibitory concentrations of MZ has been correlated with oxidative response, proteins degradation, and energy metabolism, and its main effect on yeast has been attributed to its high reactivity with thiol groups in proteins. Herein, we show that acute MZ treatments on aerobic exponentially growing yeast of wild type (BY4741) and deletion mutant strains, coupled with multiplex flow cytometry analysis, conclusively demonstrated that MZ displays the typical features of pro-oxidant activity on Saccharomyces, elevating mitochondrial ROS, and causing hyper-polarization of mitochondrial membranes leading to apoptosis. A drastic reduction of cellular viability associated with the maintenance of cell membrane integrity, as well as phosphatidyl serine externalization on yeast cells exposed to MZ, also supports an apoptotic mode of action. Moreover, abrogation of the apoptotic response in yca1 deficient mutants indicates that metacaspase-1 is involved in the programmed cell death mechanism induced by MZ in yeast.

Keywords

Dithiocarbamate Programmed cell death Yeast ROS Mitochondrial membrane potential 

Notes

Acknowledgments

The authors acknowledge Cytogene Diagnósticos Moleculares Ltda. for access to flow cytometry equipment. F. J. Scariot thanks Coordenação de Aperfeiçoamento de Pessoal de Nivel Superior for fellowship support.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10495_2016_1251_MOESM1_ESM.docx (1.7 mb)
Supplementary material 1 (DOCX 1763 kb)
10495_2016_1251_MOESM2_ESM.docx (1.5 mb)
Supplementary material 2 (DOCX 1501 kb)

References

  1. 1.
    Gullino ML, Tinivella F, Garibaldi A, Kemmitt GM, Bacci L, Sheppard B (2010) Mancozeb: past, present, and future. Plant Dis 94:1076–1087. doi: 10.1094/PDIS-94-9-1076 CrossRefGoogle Scholar
  2. 2.
    Zhou Y, Shei FS, Piccardo P, Montine TJ, Zhang J (2004) Proteosomal inhibition induced by manganese ethylene-bis-dithiocarbamate: relevance to Parkinson’s disease. Neuroscience 128:281–291. doi: 10.1016/j.neuroscience.2004.06.048 CrossRefPubMedGoogle Scholar
  3. 3.
    Corsini E, Birindelli S, Fustinoni S, De Paschale G, Mammone T, Visentin S, Galli CL, Marinovich M, Colosio C (2005) Immunomodulatory effects of the fungicide Mancozeb in agricultural workers. Toxicol Appl Pharmacol 208:178–185. doi: 10.1016/j.taap.2005.02.011 CrossRefPubMedGoogle Scholar
  4. 4.
    Kamel F, Engel LS, Gladen BC, Hoppin JA, Alavanja MC, Sandler DP (2005) Neurologic symptoms in licensed private pesticide applications in the agricultural health study. Environ Health Perspect 113:877–882. doi: 10.1289/ehp.7645 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Calviello G, Piccioni E, Boninsegna A, Tedesco B, Maggiano N, Serini S, Wolf FI, Palozza P (2006) DNA damage and apoptosis induction by the pesticide Mancozeb in rat cells: involvement of the oxidative mechanism. Toxicol Appl Pharmacol 211:87–96. doi: 10.1016/j.taap.2005.06.001 CrossRefPubMedGoogle Scholar
  6. 6.
    Domico LM, Cooper KR, Bernard LP, Zeevalk GD (2007) Reactive oxygen species generation by the ethylene-bis-dithiocarbamate (EBDC) fungicide mancozeb and its contribution to neuronal toxicity in mesencephalic cells. Neurotoxicology 28:1079–1091. doi: 10.1016/j.neuro.2007.04.008 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Srivastava AK, Ali W, Singh R, Bhui K, Tyagi S, Al-Khedhairy AA, Srivastava PK, Musarrat J, Shukla Y (2012) Mancozeb-induces genotoxicity and apoptosis in cultured human lymphocytes. Life Sci 90:815–824. doi: 10.1016/j.lfs.2011.12.013 CrossRefPubMedGoogle Scholar
  8. 8.
    Fitsanakis VA, Amarnath V, Moore JT, Montine KS, Zhang J, Montine TJ (2002) Catalysis of catechol oxidation by metal-dithiocarbamate complexes in pesticides. Free Radical Bio Med 33:1714–1723. doi: 10.1016/S0891-5849(02)01169-3 CrossRefGoogle Scholar
  9. 9.
    Xie J, Potter A, Xie W, Lynch C, Seefeldt T (2014) Evaluation of a dithiocarbamate derivative as a model of thiol oxidative stress in H9c2 rat cardiomyocytes. Free Radical Biol Med 70:214–222. doi: 10.1016/j.freeradbiomed.2014.02.022 CrossRefGoogle Scholar
  10. 10.
    Cabras P, Angioni A (2000) Pesticide residues in grapes, wine, and their processing products. J Agric Food Chem 48:967–973. doi: 10.1021/jf990727a CrossRefPubMedGoogle Scholar
  11. 11.
    Santos PM, Simões T, Sá-Correia I (2009) Insights into yeast adaptive response to the agricultural fungicide mancozeb: a toxicoproteomics approach. Proteomics 9:657–670. doi: 10.1002/pmic.200800452 CrossRefPubMedGoogle Scholar
  12. 12.
    Dias PJ, Teixeira MC, Telo JP, Sá-Correia I (2010) Insights into the mechanisms of toxicity and tolerance to the agricultural fungicide Mancozeb in yeast, as suggested by a chemogenomic approach. OMICS 14:211–227. doi: 10.1089/omi.2009.0134 CrossRefPubMedGoogle Scholar
  13. 13.
    Teixeira MC, Dia PJ, Simões T, Sá-Correia I (2008) Yeast adaptation to mancozeb involves the up-regulation of FLR1 under the coordinate control of Yap1, Rpn4, Pdr3, and Yrr1. Biochem Biophys Res 367:249–255. doi: 10.1016/j.bbrc.2007.12.056 CrossRefGoogle Scholar
  14. 14.
    Monteiro PT, Dias PJ, Ropers D, Oliveira AL, Sá-Correia I, Teixeira MC, Freitas AT (2011) Qualitative modelling and formal verification of the FLR1 gene mancozeb response in Saccharomyces cerevisiae. IET Syst Biol 5:308–316. doi: 10.1049/iet-syb.2011.0001 CrossRefPubMedGoogle Scholar
  15. 15.
    Casalone E, Bonelli E, Polsinelli M (2010) Effects of Mancozeb and other dithiocarbamate fungicides on Saccharomyces cerevisiae: the role of mitochondrial petite mutants in dithiocarbamate tolerance. Folia Microbiol 55:593–597. doi: 10.1007/s12223-010-0095-5 CrossRefGoogle Scholar
  16. 16.
    Delobel P, Tesnière C (2014) A simple FCM method to avoid misinterpretation of Saccharomyces cerevisiae cell cycle assessment between G0 and sub-G1. PLoS One 9:e84645. doi: 10.1371/journal.pone.0084645 CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Carmona-Gutierrez D, Eisenberg T, Büttner S, Meisinger C, Kroemer G, Madeo F (2010) Apoptosis in yeast: triggers, pathways, subroutines. Cell Death Differ 17:763–773. doi: 10.1038/cdd.2009.219 CrossRefPubMedGoogle Scholar
  18. 18.
    Wloch-Salomon DM, Bem AE (2012) Types of cell death and methods of their detection in yeast Saccharomyces cerevisiae. J Appl Microbiol 114:287–298. doi: 10.1111/jam.12024 CrossRefGoogle Scholar
  19. 19.
    Perrone GG, Tan SX, Dawes IW (2008) Reactive oxygen species and yeast apoptosis. Biochem Biophys Acta 1783:1354–1368. doi: 10.1016/j.bbamcr.2008.01.023 CrossRefPubMedGoogle Scholar
  20. 20.
    Farrugia G, Balzan R (2012) Oxidative stress and programmed cell death in yeast. Front Oncol 2:1–21. doi: 10.3389/fonc.2012.00064 CrossRefGoogle Scholar
  21. 21.
    Bussche JV, Soares EV (2011) Lead induces oxidative stress and phenotypic markers of apoptosis in Saccharomyces. Appl Microbiol Biotechnol 90:679–687. doi: 10.1007/s00253-010-3056-7 CrossRefPubMedGoogle Scholar
  22. 22.
    Madeo F, Frohlich E, Frohlich KU (1997) A yeast mutant showing diagnostic markers of early and late apoptosis. J Cell Biol 139:729–734. doi: 10.1083/jcb.139.3.729 CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Lefevre S, Sliwa D, Auchère F, Brossas C, Ruckenstuhl C, Boggetto N, Lesuisse E, Madeo F, Camadro JM, Santos R (2012) The yeast metacaspase is implicated in oxidative stress response in frataxin-deficient cells. FEBS Lett 586:143–148. doi: 10.1016/j.febslet.2011.12.002 CrossRefPubMedGoogle Scholar
  24. 24.
    Zdralevic M, Guaragnella N, Antonacci L, Marra E, Giannattasio S (2012) Yeast as a tool to study signaling pathways in mitochondrial stress response and cytoprotection. ScientificWorldJournal. doi: 10.1100/2012/912147 PubMedPubMedCentralGoogle Scholar
  25. 25.
    Pozniakovsky AI, Knorre DA, Markova OV, Hyman AA, Skulachev VP, Severin F (2005) Role of mitochondria in the pheromone- and amiodarone-induced programmed death of yeast. J Cell Biol 168:257–269. doi: 10.1083/jcb.200408145 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Nargund AM, Avery SV, Houghton JE (2008) Cadmium induces a heterogeneous and caspase-dependent apoptotic response in Saccharomyces cerevisiae. Apoptosis 13:811–821. doi: 10.1007/s10495-008-0215-8 CrossRefPubMedGoogle Scholar
  27. 27.
    Pereira C, Silva RD, Saraiva L, Johansson B, Sousa MJ, Côrte-Real M (2008) Mitochondria-dependent apoptosis in yeast. Biochim Biophys Acta 1783:1286–1302. doi: 10.1016/j.bbamcr.2008.03.010 CrossRefPubMedGoogle Scholar
  28. 28.
    Pagani MA, Casamayor A, Serrano R, Atrian S, Ariño J (2007) Disruption of iron homeostasis in Saccharomyces cerevisiae by high zinc levels: a genome-wide study. Mol Microbiol 65:521–537. doi: 10.1016/j.bbamcr.2008.03.010 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • F. J. Scariot
    • 1
  • L. M. Jahn
    • 1
  • J. P. Maianti
    • 2
  • A. P. L. Delamare
    • 1
  • S. Echeverrigaray
    • 1
    • 3
    Email author
  1. 1.Laboratory of Applied Microbiology, Institute of BiotechnologyUniversity of Caxias do SulCaxias do SulBrazil
  2. 2.Department of Chemistry and Chemical BiologyHarvard UniversityCambridgeUSA
  3. 3.Cytogene Diagnósticos Moleculares Ltda.LageadoBrazil

Personalised recommendations