Yeast polyubiquitin gene UBI4 deficiency leads to early induction of apoptosis and shortened replicative lifespan

  • Wei Zhao
  • Tao Zhou
  • Hua-Zhen Zheng
  • Kun-Pei Qiu
  • Hong-Jing Cui
  • Hui Yu
  • Xin-Guang Liu
Original Paper

Abstract

Ubiquitin is a 76-amino acid protein that is highly conserved among higher and lower eukaryotes. The polyubiquitin gene UBI4 encodes a unique precursor protein that contains five ubiquitin repeats organized in a head-to-tail arrangement. Although the involvement of the yeast polyubiquitin gene UBI4 in the stress response was reported long ago, there are no reports regarding the underlying mechanism of this involvement. In this study, we used UBI4-deletion and UBI4-overexpressing yeast strains as models to explore the potential mechanism by which UBI4 protects yeast cells against paraquat-induced oxidative stress. Here, we show that ubi4Δ cells exhibit oxidative stress, an apoptotic phenotype, and a decreased replicative lifespan. Additionally, the reduced resistance of ubi4Δ cells to paraquat that was observed in this study was rescued by overexpression of either the catalase or the mitochondrial superoxide dismutase SOD2. We also demonstrated that only SOD2 overexpression restored the replicative lifespan of ubi4Δ cells. In contrast to the case of ubi4Δ cells, UBI4 overexpression in wild-type yeast increases the yeast’s resistance to paraquat, and this overexpression is associated with large pools of expressed ubiquitin and increased levels of ubiquitinated proteins. Collectively, these findings highlight the role of the polyubiquitin gene UBI4 in apoptosis and implicate UBI4 as a modulator of the replicative lifespan.

Keywords

Yeast UBI4 Apoptosis Replicative lifespan 

Notes

Acknowledgements

We are grateful to Brian K. Kennedy (Buck Institute), Matt Kaeberlein, and Brian M. Wasko (University of Washington) for technical assistance.

Supplementary material

12192_2017_860_MOESM1_ESM.docx (119 kb)
Suppl. Fig 1 (DOCX 119 kb).
12192_2017_860_MOESM2_ESM.docx (88 kb)
Suppl. Fig 2 (DOCX 87 kb).
12192_2017_860_MOESM3_ESM.xls (28 kb)
Table S1 (XLS 27 kb).
12192_2017_860_MOESM4_ESM.xls (26 kb)
Table S2 (XLS 25 kb).

References

  1. Aguilar RC, Wendland B (2003) Ubiquitin: not just for proteasomes anymore. Curr Opin Cell Biol 15(2):184–190CrossRefPubMedGoogle Scholar
  2. Altomare RE, Greenfield PF, Kittrell JR (1974) Letter: inactivation of immobilized fungal catalase by hydrogen peroxide. Biotechnol Bioeng 16(12):1675–1680CrossRefPubMedGoogle Scholar
  3. Ayer A, Gourlay CW, Dawes IW (2014) Cellular redox homeostasis, reactive oxygen species and replicative ageing in Saccharomyces cerevisiae. FEMS Yeast Res 14(1):60–72CrossRefPubMedGoogle Scholar
  4. Baudin A, Ozier-Kalogeropoulos O, Denouel A, Lacroute F, Cullin C (1993) A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae. Nucleic Acids Res 21(14):3329–3330CrossRefPubMedPubMedCentralGoogle Scholar
  5. Breitenbach M (2005) Apoptosis, ageing and redox homeostasis in yeasts. FEMS Yeast Res 5(12):1191–1192CrossRefPubMedGoogle Scholar
  6. Buckley SM, Aranda-Orgilles B, Strikoudis A, Apostolou E, Loizou E, Moran-Crusio K, Farnsworth CL, Koller AA, Dasgupta R, Silva JC, Stadtfeld M, Hochedlinger K, Chen EI, Aifantis I (2012) Regulation of pluripotency and cellular reprogramming by the ubiquitin-proteasome system. Cell Stem Cell 11(6):783–798CrossRefPubMedPubMedCentralGoogle Scholar
  7. Chen SR, Dunigan DD, Dickman MB (2003) Bcl-2 family members inhibit oxidative stress-induced programmed cell death in Saccharomyces cerevisiae. Free Radic Biol Med 34(10):1315–1325CrossRefPubMedGoogle Scholar
  8. Chen Y, Piper PW (1995) Consequences of the overexpression of ubiquitin in yeast: elevated tolerances of osmostress, ethanol and canavanine, yet reduced tolerances of cadmium, arsenite and paromomycin. Biochim Biophys Acta 1268(1):59–64CrossRefPubMedGoogle Scholar
  9. Choi SI, Kim TI, Kim KS, Kim BY, Ahn SY, Cho HJ, Lee HK, Cho HS, Kim EK (2009) Decreased catalase expression and increased susceptibility to oxidative stress in primary cultured corneal fibroblasts from patients with granular corneal dystrophy type II. Am J Pathol 175(1):248–261CrossRefPubMedPubMedCentralGoogle Scholar
  10. Cocheme HM, Murphy MP (2008) Complex I is the major site of mitochondrial superoxide production by paraquat. J Biol Chem 283(4):1786–1798CrossRefPubMedGoogle Scholar
  11. Cohen G, Rapatz W, Ruis H (1988) Sequence of the Saccharomyces cerevisiae CTA1 gene and amino acid sequence of catalase A derived from it. Eur J Biochem 176(1):159–163CrossRefPubMedGoogle Scholar
  12. Delaney JR, Ahmed U, Chou A, Sim S, Carr D, Murakami CJ, Schleit J, Sutphin GL, An EH, Castanza A, Fletcher M, Higgins S, Jelic M, Klum S, Muller B, Peng ZJ, Rai D, Ros V, Singh M, Wende HV, Kennedy BK, Kaeberlein M (2013) Stress profiling of longevity mutants identifies Afg3 as a mitochondrial determinant of cytoplasmic mRNA translation and aging. Aging Cell 12(1):156–166CrossRefPubMedGoogle Scholar
  13. Fedoseeva IV, Pyatrikas DV, Stepanov AV, Fedyaeva AV, Varakina NN, Rusaleva TM, Borovskii GB, Rikhvanov EG (2017) The role of flavin-containing enzymes in mitochondrial membrane hyperpolarization and ROS production in respiring Saccharomyces cerevisiae cells under heat-shock conditions. Sci Rep 7(1):2586CrossRefPubMedPubMedCentralGoogle Scholar
  14. Finley D, Chau V (1991) Ubiquitination. Annu Rev Cell Biol 7:25–69CrossRefPubMedGoogle Scholar
  15. Finley D, Ozkaynak E, Varshavsky A (1987) The yeast polyubiquitin gene is essential for resistance to high temperatures, starvation, and other stresses. Cell 48(6):1035–1046CrossRefPubMedGoogle Scholar
  16. Fraser J, Luu HA, Neculcea J, Thomas DY, Storms RK (1991) Ubiquitin gene expression: response to environmental changes. Curr Genet 20(1–2):17–23CrossRefPubMedGoogle Scholar
  17. Frohlich KU, Madeo F (2001) Apoptosis in yeast: a new model for aging research. Exp Gerontol 37(1):27–31CrossRefPubMedGoogle Scholar
  18. Gao C, Xing D, Li L, Zhang L (2008) Implication of reactive oxygen species and mitochondrial dysfunction in the early stages of plant programmed cell death induced by ultraviolet-C overexposure. Planta 227(4):755–767CrossRefPubMedGoogle Scholar
  19. Goldberg AL (2003) Protein degradation and protection against misfolded or damaged proteins. Nature 426(6968):895–899CrossRefPubMedGoogle Scholar
  20. Hartig A, Ruis H (1986) Nucleotide sequence of the Saccharomyces cerevisiae CTT1 gene and deduced amino-acid sequence of yeast catalase T. Eur J Biochem 160(3):487–490CrossRefPubMedGoogle Scholar
  21. Herker E, Jungwirth H, Lehmann KA, Maldener C, Frohlich KU, Wissing S, Buttner S, Fehr M, Sigrist S, Madeo F (2004) Chronological aging leads to apoptosis in yeast. J Cell Biol 164(4):501–507CrossRefPubMedPubMedCentralGoogle Scholar
  22. Huang CL, Chao CC, Lee YC, MK L, Cheng JJ, Yang YC, Wang VC, Chang WC, Huang NK (2016) Paraquat induces cell death through impairing mitochondrial membrane permeability. Mol Neurobiol 53(4):2169–2188CrossRefPubMedGoogle Scholar
  23. Iwai K, Kondo T, Watanabe M, Yabu T, Kitano T, Taguchi Y, Umehara H, Takahashi A, Uchiyama T, Okazaki T (2003) Ceramide increases oxidative damage due to inhibition of catalase by caspase-3-dependent proteolysis in HL-60 cell apoptosis. J Biol Chem 278(11):9813–9822CrossRefPubMedGoogle Scholar
  24. Jackson SP, Durocher D (2013) Regulation of DNA damage responses by ubiquitin and SUMO. Mol Cell 49(5):795–807CrossRefPubMedGoogle Scholar
  25. Jamieson DJ (1998) Oxidative stress responses of the yeast Saccharomyces cerevisiae. Yeast 14(16):1511–1527CrossRefPubMedGoogle Scholar
  26. Kim JM, Bae HR, Park BS, Lee JM, Ahn HB, Rho JH, Yoo KW, Park WC, Rho SH, Yoon HS, Yoo YH (2003) Early mitochondrial hyperpolarization and intracellular alkalinization in lactacystin-induced apoptosis of retinal pigment epithelial cells. J Pharmacol Exp Ther 305(2):474–481CrossRefPubMedGoogle Scholar
  27. Laun P, Buttner S, Rinnerthaler M, Burhans WC, Breitenbach M (2012) Yeast aging and apoptosis. Subcell Biochem 57:207–232CrossRefPubMedGoogle Scholar
  28. Laun P, Heeren G, Rinnerthaler M, Rid R, Kossler S, Koller L, Breitenbach M (2008) Senescence and apoptosis in yeast mother cell-specific aging and in higher cells: a short review. Biochim Biophys Acta 1783(7):1328–1334CrossRefPubMedGoogle Scholar
  29. Ligr M, Velten I, Frohlich E, Madeo F, Ledig M, Frohlich KU, Wolf DH, Hilt W (2001) The proteasomal substrate Stm1 participates in apoptosis-like cell death in yeast. Mol Biol Cell 12(8):2422–2432CrossRefPubMedPubMedCentralGoogle Scholar
  30. Lin MT, Beal MF (2006) Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature 443(7113):787–795CrossRefPubMedGoogle Scholar
  31. Ludovico P, Sansonetty F, Corte-Real M (2001) Assessment of mitochondrial membrane potential in yeast cell populations by flow cytometry. Microbiology 147(Pt 12):3335–3343CrossRefPubMedGoogle Scholar
  32. Martin SJ, Reutelingsperger CP, McGahon AJ, Rader JA, van Schie RC, LaFace DM, Green DR (1995) Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med 182(5):1545–1556CrossRefPubMedGoogle Scholar
  33. Myeku N, Figueiredo-Pereira ME (2011) Dynamics of the degradation of ubiquitinated proteins by proteasomes and autophagy: association with sequestosome 1/p62. J Biol Chem 286(25):22426–22440CrossRefPubMedPubMedCentralGoogle Scholar
  34. Navarro-Yepes J, Anandhan A, Bradley E, Bohovych I, Yarabe B, de Jong A, Ovaa H, Zhou Y, Khalimonchuk O, Quintanilla-Vega B, Franco R (2016) Inhibition of protein ubiquitination by paraquat and 1-methyl-4-phenylpyridinium impairs ubiquitin-dependent protein degradation pathways. Mol Neurobiol 53(8):5229–5251CrossRefPubMedGoogle Scholar
  35. Ozkaynak E, Finley D, Solomon MJ, Varshavsky A (1987) The yeast ubiquitin genes: a family of natural gene fusions. EMBO J 6(5):1429–1439PubMedPubMedCentralGoogle Scholar
  36. Pyatrikas DV, Fedoseeva IV, Varakina NN, Rusaleva TM, Stepanov AV, Fedyaeva AV, Borovskii GB, Rikhvanov EG (2015) Relation between cell death progression, reactive oxygen species production and mitochondrial membrane potential in fermenting Saccharomyces cerevisiae cells under heat-shock conditions. FEMS Microbiol Lett 362(12):82CrossRefGoogle Scholar
  37. Rajendra E, Oestergaard VH, Langevin F, Wang M, Dornan GL, Patel KJ, Passmore LA (2014) The genetic and biochemical basis of FANCD2 monoubiquitination. Mol Cell 54(5):858–869CrossRefPubMedPubMedCentralGoogle Scholar
  38. Rona G, Herdeiro R, Mathias CJ, Torres FA, Pereira MD, Eleutherio E (2015) CTT1 overexpression increases life span of calorie-restricted Saccharomyces cerevisiae deficient in Sod1. Biogerontology 16(3):343–351CrossRefPubMedGoogle Scholar
  39. Schagger H (2006) Tricine-SDS-PAGE. Nat Protoc 1(1):16–22CrossRefPubMedGoogle Scholar
  40. Shang F, Taylor A (2011) Ubiquitin-proteasome pathway and cellular responses to oxidative stress. Free Radic Biol Med 51(1):5–16CrossRefPubMedPubMedCentralGoogle Scholar
  41. Stearns T, Ma H, Botstein D (1990) Manipulating yeast genome using plasmid vectors. Methods Enzymol 185:280–297CrossRefPubMedGoogle Scholar
  42. Steffen KK, Kennedy BK, Kaeberlein M (2009) Measuring replicative life span in the budding yeast. J Vis Exp 28:1209Google Scholar
  43. Suntres ZE (2002) Role of antioxidants in paraquat toxicity. Toxicology 180(1):65–77CrossRefPubMedGoogle Scholar
  44. Watt R, Piper PW (1997) UBI4, the polyubiquitin gene of Saccharomyces cerevisiae, is a heat shock gene that is also subject to catabolite derepression control. Mol Gen Genet 253(4):439–447CrossRefPubMedGoogle Scholar
  45. Wing SS, Haas AL, Goldberg AL (1995) Increase in ubiquitin-protein conjugates concomitant with the increase in proteolysis in rat skeletal muscle during starvation and atrophy denervation. Biochem J 307(Pt 3):639–645CrossRefPubMedPubMedCentralGoogle Scholar
  46. Yang W, Chen L, Ding Y, Zhuang X, Kang UJ (2007) Paraquat induces dopaminergic dysfunction and proteasome impairment in DJ-1-deficient mice. Hum Mol Genet 16(23):2900–2910CrossRefPubMedGoogle Scholar
  47. Zhang D, Ren L, Chen GQ, Zhang J, Reed BM, Shen XH (2015) ROS-induced oxidative stress and apoptosis-like event directly affect the cell viability of cryopreserved embryogenic callus in Agapanthus praecox. Plant Cell Rep 34(9):1499–1513CrossRefPubMedGoogle Scholar
  48. Zhao W, Fang BX, Niu YJ, Liu YN, Liu B, Peng Q, Li JB, Wasko BM, Delaney JR, Kennedy BK, Suh Y, Zhou ZJ, Kaeberlein M, Liu XG (2014) Nar1 deficiency results in shortened lifespan and sensitivity to paraquat that is rescued by increased expression of mitochondrial superoxide dismutase. Mech Ageing Dev 138:53–58CrossRefPubMedGoogle Scholar

Copyright information

© Cell Stress Society International 2017

Authors and Affiliations

  • Wei Zhao
    • 1
    • 2
  • Tao Zhou
    • 1
    • 2
  • Hua-Zhen Zheng
    • 1
    • 3
  • Kun-Pei Qiu
    • 1
    • 2
  • Hong-Jing Cui
    • 1
    • 2
  • Hui Yu
    • 1
    • 2
  • Xin-Guang Liu
    • 1
    • 2
    • 4
  1. 1.Institute of Aging ResearchGuangdong Medical UniversityDongguanChina
  2. 2.Guangdong Provincial Key Laboratory of Medical Molecular DiagnosticsDongguanChina
  3. 3.Department of Clinical LaboratoryThe First People’s Hospital of FoshanFoshanChina
  4. 4.Institute of Biochemistry and Molecular BiologyGuangdong Medical UniversityDongguanChina

Personalised recommendations