Yeast polyubiquitin gene UBI4 deficiency leads to early induction of apoptosis and shortened replicative lifespan
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.
KeywordsYeast UBI4 Apoptosis Replicative lifespan
We are grateful to Brian K. Kennedy (Buck Institute), Matt Kaeberlein, and Brian M. Wasko (University of Washington) for technical assistance.
This work was supported by the China National Natural Science Foundation (31101051, 81671399), the Ordinary University Innovation Team Construction Project of Guangdong Province (2015KCXTD022), the Unique Innovative Projects in Ordinary University of Guangdong Province (2015KTSCX049), and the Guangdong Medical Research Foundation (A2016257).
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- 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
- Steffen KK, Kennedy BK, Kaeberlein M (2009) Measuring replicative life span in the budding yeast. J Vis Exp 28:1209Google Scholar
- 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