14 research outputs found
Different Toxicity Mechanisms for Citrinin and Ochratoxin A Revealed by Transcriptomic Analysis in Yeast
[EN] Citrinin (CIT) and ochratoxin A (OTA) are important mycotoxins, which frequently co-contaminate foodstuff. In order to assess the toxicologic threat posed by the two mycotoxins separately or in combination, their biological effects were studied here using genomic transcription profiling and specific live cell gene expression reporters in yeast cells. Both CIT and OTA cause highly transient transcriptional activation of different stress genes, which is greatly enhanced by the disruption of the multidrug exporter Pdr5. Therefore, we performed genome-wide transcription profiling experiments with the pdr5 mutant in response to acute CIT, OTA, or combined CIT/OTA exposure. We found that CIT and OTA activate divergent and largely nonoverlapping gene sets in yeast. CIT mainly caused the rapid induction of antioxidant and drug extrusion-related gene functions, while OTA mainly deregulated developmental genes related with yeast sporulation and sexual reproduction, having only a minor effect on the antioxidant response. The simultaneous exposure to CIT and OTA gave rise to a genomic response, which combined the specific features of the separated mycotoxin treatments. The application of stress-specific mutants and reporter gene fusions further confirmed that both mycotoxins have divergent biological effects in cells. Our results indicate that CIT exposure causes a strong oxidative stress, which triggers a massive transcriptional antioxidant and drug extrusion response, while OTA mainly deregulates developmental genes and only marginally induces the antioxidant defense.We thank Lorena Latorre and Javier Forment for their help with the microarray experiments and data analysis. This work was funded only in the initial phase by a grant from Ministerio de EconomĂa y Competitividad (BFU2011-23326). We thank the Fond for Open Access Publication from Consejo Superior de Investigaciones CientĂficas (CSIC) for supporting publication costs of this article.Vanacloig-PedrĂłs, E.; Proft, MH.; Pascual-Ahuir Giner, MD. (2016). Different Toxicity Mechanisms for Citrinin and Ochratoxin A Revealed by Transcriptomic Analysis in Yeast. Toxins. 8(10):1-20. https://doi.org/10.3390/toxins8100273S120810Bennett, J. W., & Klich, M. (2003). Mycotoxins. Clinical Microbiology Reviews, 16(3), 497-516. doi:10.1128/cmr.16.3.497-516.2003MarroquĂn-Cardona, A. G., Johnson, N. M., Phillips, T. D., & Hayes, A. W. (2014). Mycotoxins in a changing global environment – A review. Food and Chemical Toxicology, 69, 220-230. doi:10.1016/j.fct.2014.04.025Moretti, A., Susca, A., MulĂ©, G., Logrieco, A. F., & Proctor, R. H. (2013). Molecular biodiversity of mycotoxigenic fungi that threaten food safety. International Journal of Food Microbiology, 167(1), 57-66. doi:10.1016/j.ijfoodmicro.2013.06.033Wu, F., Groopman, J. D., & Pestka, J. J. (2014). Public Health Impacts of Foodborne Mycotoxins. Annual Review of Food Science and Technology, 5(1), 351-372. doi:10.1146/annurev-food-030713-092431Möbius, N., & Hertweck, C. (2009). Fungal phytotoxins as mediators of virulence. Current Opinion in Plant Biology, 12(4), 390-398. doi:10.1016/j.pbi.2009.06.004Doi, K., & Uetsuka, K. (2014). Mechanisms of Mycotoxin-induced Dermal Toxicity and Tumorigenesis Through Oxidative Stress-related Pathways. Journal of Toxicologic Pathology, 27(1), 1-10. doi:10.1293/tox.2013-0062Escrivá, L., Font, G., & Manyes, L. (2015). In vivo toxicity studies of fusarium mycotoxins in the last decade: A review. Food and Chemical Toxicology, 78, 185-206. doi:10.1016/j.fct.2015.02.005Vettorazzi, A., González-Peñas, E., & de Cerain, A. L. (2014). Ochratoxin A kinetics: A review of analytical methods and studies in rat model. Food and Chemical Toxicology, 72, 273-288. doi:10.1016/j.fct.2014.07.020Wang, Y., Wang, L., Liu, F., Wang, Q., Selvaraj, J., Xing, F., … Liu, Y. (2016). Ochratoxin A Producing Fungi, Biosynthetic Pathway and Regulatory Mechanisms. Toxins, 8(3), 83. doi:10.3390/toxins8030083KĹ‘szegi, T., & PoĂłr, M. (2016). Ochratoxin A: Molecular Interactions, Mechanisms of Toxicity and Prevention at the Molecular Level. Toxins, 8(4), 111. doi:10.3390/toxins8040111Faucet, V., Pfohl-Leszkowicz, A., Dai, J., Castegnaro, M., & Manderville, R. A. (2004). Evidence for Covalent DNA Adduction by Ochratoxin A following Chronic Exposure to Rat and Subacute Exposure to Pig. Chemical Research in Toxicology, 17(9), 1289-1296. doi:10.1021/tx049877sMantle, P. G., Faucet-Marquis, V., Manderville, R. A., Squillaci, B., & Pfohl-Leszkowicz, A. (2010). Structures of Covalent Adducts between DNA and Ochratoxin A: A New Factor in Debate about Genotoxicity and Human Risk Assessment. Chemical Research in Toxicology, 23(1), 89-98. doi:10.1021/tx900295aPfohl-Leszkowicz, A., & Manderville, R. A. (2011). An Update on Direct Genotoxicity as a Molecular Mechanism of Ochratoxin A Carcinogenicity. Chemical Research in Toxicology, 25(2), 252-262. doi:10.1021/tx200430fRahimtula, A. D., BĂ©rĂ©ziat, J.-C., Bussacchini-Griot, V., & Bartsch, H. (1988). Lipid peroxidation as a possible cause of ochratoxin a toxicity. Biochemical Pharmacology, 37(23), 4469-4477. doi:10.1016/0006-2952(88)90662-4Sorrenti, V., Di Giacomo, C., Acquaviva, R., Barbagallo, I., Bognanno, M., & Galvano, F. (2013). Toxicity of Ochratoxin A and Its Modulation by Antioxidants: A Review. Toxins, 5(10), 1742-1766. doi:10.3390/toxins5101742BRAGULAT, M., MARTINEZ, E., CASTELLA, G., & CABANES, F. (2008). Ochratoxin A and citrinin producing species of the genus Penicillium from feedstuffs. International Journal of Food Microbiology, 126(1-2), 43-48. doi:10.1016/j.ijfoodmicro.2008.04.034Vrabcheva, T., Usleber, E., Dietrich, R., & Märtlbauer, E. (2000). Co-occurrence of Ochratoxin A and Citrinin in Cereals from Bulgarian Villages with a History of Balkan Endemic Nephropathy. Journal of Agricultural and Food Chemistry, 48(6), 2483-2488. doi:10.1021/jf990891yOstry, V., Malir, F., & Ruprich, J. (2013). Producers and Important Dietary Sources of Ochratoxin A and Citrinin. Toxins, 5(9), 1574-1586. doi:10.3390/toxins5091574Schmidt-Heydt, M., Graf, E., Stoll, D., & Geisen, R. (2012). The biosynthesis of ochratoxin A by Penicillium as one mechanism for adaptation to NaCl rich foods. Food Microbiology, 29(2), 233-241. doi:10.1016/j.fm.2011.08.003Schmidt-Heydt, M., Stoll, D., SchĂĽtz, P., & Geisen, R. (2015). Oxidative stress induces the biosynthesis of citrinin by Penicillium verrucosum at the expense of ochratoxin. International Journal of Food Microbiology, 192, 1-6. doi:10.1016/j.ijfoodmicro.2014.09.008Stoll, D., Schmidt-Heydt, M., & Geisen, R. (2013). Differences in the Regulation of Ochratoxin A by the HOG Pathway in Penicillium and Aspergillus in Response to High Osmolar Environments. Toxins, 5(7), 1282-1298. doi:10.3390/toxins5071282Flajs, D., & Peraica, M. (2009). Toxicological Properties of Citrinin. Archives of Industrial Hygiene and Toxicology, 60(4), 457-464. doi:10.2478/10004-1254-60-2009-1992Bouslimi, A., Ouannes, Z., Golli, E. E., Bouaziz, C., Hassen, W., & Bacha, H. (2008). Cytotoxicity and Oxidative Damage in Kidney Cells Exposed to the Mycotoxins Ochratoxin A and Citrinin: Individual and Combined Effects. Toxicology Mechanisms and Methods, 18(4), 341-349. doi:10.1080/15376510701556682Chan, W.-H. (2007). Citrinin induces apoptosis via a mitochondria-dependent pathway and inhibition of survival signals in embryonic stem cells, and causes developmental injury in blastocysts. Biochemical Journal, 404(2), 317-326. doi:10.1042/bj20061875Kumar, M., Dwivedi, P., Sharma, A. K., Sankar, M., Patil, R. D., & Singh, N. D. (2012). Apoptosis and lipid peroxidation in ochratoxin A- and citrinin-induced nephrotoxicity in rabbits. Toxicology and Industrial Health, 30(1), 90-98. doi:10.1177/0748233712452598Kumar, R., Dwivedi, P. D., Dhawan, A., Das, M., & Ansari, K. M. (2011). Citrinin-Generated Reactive Oxygen Species Cause Cell Cycle Arrest Leading to Apoptosis via the Intrinsic Mitochondrial Pathway in Mouse Skin. Toxicological Sciences, 122(2), 557-566. doi:10.1093/toxsci/kfr143MátĂ©, G., Gazdag, Z., Mike, N., Papp, G., PĂłcsi, I., & Pesti, M. (2014). Regulation of oxidative stress-induced cytotoxic processes of citrinin in the fission yeast Schizosaccharomyces pombe. Toxicon, 90, 155-166. doi:10.1016/j.toxicon.2014.08.005Pascual-Ahuir, A., Vanacloig-Pedros, E., & Proft, M. (2014). Toxicity Mechanisms of the Food Contaminant Citrinin: Application of a Quantitative Yeast Model. Nutrients, 6(5), 2077-2087. doi:10.3390/nu6052077Ribeiro, S. M. R., Chagas, G. M., Campello, A. P., & Kluppel, M. L. W. (1997). Mechanism of citrinin-induced dysfunction of mitochondria. V. Effect on the homeostasis of the reactive oxygen species. Cell Biochemistry and Function, 15(3), 203-209. doi:10.1002/(sici)1099-0844(199709)15:33.0.co;2-jSingh, N. D., Sharma, A. K., Dwivedi, P., Leishangthem, G. D., Rahman, S., Reddy, J., & Kumar, M. (2013). Effect of feeding graded doses of citrinin on apoptosis and oxidative stress in male Wistar rats through the F1generation. Toxicology and Industrial Health, 32(3), 385-397. doi:10.1177/0748233713500836Yu, F.-Y., Liao, Y.-C., Chang, C.-H., & Liu, B.-H. (2006). Citrinin induces apoptosis in HL-60 cells via activation of the mitochondrial pathway. Toxicology Letters, 161(2), 143-151. doi:10.1016/j.toxlet.2005.08.009Föllmann, W., Behm, C., & Degen, G. H. (2014). Toxicity of the mycotoxin citrinin and its metabolite dihydrocitrinone and of mixtures of citrinin and ochratoxin A in vitro. Archives of Toxicology, 88(5), 1097-1107. doi:10.1007/s00204-014-1216-8Klarić, M., Rašić, D., & Peraica, M. (2013). Deleterious Effects of Mycotoxin Combinations Involving Ochratoxin A. Toxins, 5(11), 1965-1987. doi:10.3390/toxins5111965Afshari, C. A., Hamadeh, H. K., & Bushel, P. R. (2010). The Evolution of Bioinformatics in Toxicology: Advancing Toxicogenomics. Toxicological Sciences, 120(Supplement 1), S225-S237. doi:10.1093/toxsci/kfq373Yasokawa, D., & Iwahashi, H. (2010). Toxicogenomics using yeast DNA microarrays. Journal of Bioscience and Bioengineering, 110(5), 511-522. doi:10.1016/j.jbiosc.2010.06.003Arbillaga, L., Azqueta, A., van Delft, J. H. M., & LĂłpez de Cerain, A. (2007). In vitro gene expression data supporting a DNA non-reactive genotoxic mechanism for ochratoxin A. Toxicology and Applied Pharmacology, 220(2), 216-224. doi:10.1016/j.taap.2007.01.008Hibi, D., Kijima, A., Kuroda, K., Suzuki, Y., Ishii, Y., Jin, M., … Umemura, T. (2013). Molecular mechanisms underlying ochratoxin A-induced genotoxicity: global gene expression analysis suggests induction of DNA double-strand breaks and cell cycle progression. The Journal of Toxicological Sciences, 38(1), 57-69. doi:10.2131/jts.38.57Marin-Kuan, M., Nestler, S., Verguet, C., Bezençon, C., Piguet, D., Mansourian, R., … Schilter, B. (2005). A Toxicogenomics Approach to Identify New Plausible Epigenetic Mechanisms of Ochratoxin A Carcinogenicity in Rat. Toxicological Sciences, 89(1), 120-134. doi:10.1093/toxsci/kfj017Vettorazzi, A., van Delft, J., & LĂłpez de Cerain, A. (2013). A review on ochratoxin A transcriptomic studies. Food and Chemical Toxicology, 59, 766-783. doi:10.1016/j.fct.2013.05.043Iwahashi, H., Kitagawa, E., Suzuki, Y., Ueda, Y., Ishizawa, Y., Nobumasa, H., … Iwahashi, Y. (2007). Evaluation of toxicity of the mycotoxin citrinin using yeast ORF DNA microarray and Oligo DNA microarray. BMC Genomics, 8(1), 95. doi:10.1186/1471-2164-8-95Toone, W. M., Morgan, B. A., & Jones, N. (2001). Redox control of AP-1-like factors in yeast and beyond. Oncogene, 20(19), 2336-2346. doi:10.1038/sj.onc.1204384Luo, Y., Wang, J., Liu, B., Wang, Z., Yuan, Y., & Yue, T. (2015). Effect of Yeast Cell Morphology, Cell Wall Physical Structure and Chemical Composition on Patulin Adsorption. PLOS ONE, 10(8), e0136045. doi:10.1371/journal.pone.0136045Piotrowska, M., & Masek, A. (2015). Saccharomyces Cerevisiae Cell Wall Components as Tools for Ochratoxin A Decontamination. Toxins, 7(4), 1151-1162. doi:10.3390/toxins7041151Jungwirth, H., & Kuchler, K. (2005). Yeast ABC transporters - A tale of sex, stress, drugs and aging. FEBS Letters, 580(4), 1131-1138. doi:10.1016/j.febslet.2005.12.050Prasad, R., & Goffeau, A. (2012). Yeast ATP-Binding Cassette Transporters Conferring Multidrug Resistance. Annual Review of Microbiology, 66(1), 39-63. doi:10.1146/annurev-micro-092611-150111Thakur, J. K., Arthanari, H., Yang, F., Pan, S.-J., Fan, X., Breger, J., … Näär, A. M. (2008). A nuclear receptor-like pathway regulating multidrug resistance in fungi. Nature, 452(7187), 604-609. doi:10.1038/nature06836Chen, C.-C., & Chan, W.-H. (2009). Inhibition of Citrinin-Induced Apoptotic Biochemical Signaling in Human Hepatoma G2 Cells by Resveratrol. International Journal of Molecular Sciences, 10(8), 3338-3357. doi:10.3390/ijms10083338Gayathri, L., Dhivya, R., Dhanasekaran, D., Periasamy, V. S., Alshatwi, A. A., & Akbarsha, M. A. (2015). Hepatotoxic effect of ochratoxin A and citrinin, alone and in combination, and protective effect of vitamin E: In vitro study in HepG2 cell. Food and Chemical Toxicology, 83, 151-163. doi:10.1016/j.fct.2015.06.009ALEO, M. (1991). The role of altered mitochondrial function in citrinin-induced toxicity to rat renal proximal tubule suspensions*1. Toxicology and Applied Pharmacology, 109(3), 455-463. doi:10.1016/0041-008x(91)90008-3Qi, X., Yu, T., Zhu, L., Gao, J., He, X., Huang, K., … Xu, W. (2014). Ochratoxin A induces rat renal carcinogenicity with limited induction of oxidative stress responses. Toxicology and Applied Pharmacology, 280(3), 543-549. doi:10.1016/j.taap.2014.08.030Taniai, E., Yafune, A., Nakajima, M., Hayashi, S.-M., Nakane, F., Itahashi, M., & Shibutani, M. (2014). Ochratoxin A induces karyomegaly and cell cycle aberrations in renal tubular cells without relation to induction of oxidative stress responses in rats. Toxicology Letters, 224(1), 64-72. doi:10.1016/j.toxlet.2013.10.001Govin, J., & Berger, S. L. (2009). Genome reprogramming during sporulation. The International Journal of Developmental Biology, 53(2-3), 425-432. doi:10.1387/ijdb.082687jgWinter, E. (2012). The Sum1/Ndt80 Transcriptional Switch and Commitment to Meiosis in Saccharomyces cerevisiae. Microbiology and Molecular Biology Reviews, 76(1), 1-15. doi:10.1128/mmbr.05010-11Grunstein, M., & Gasser, S. M. (2013). Epigenetics in Saccharomyces cerevisiae. Cold Spring Harbor Perspectives in Biology, 5(7), a017491-a017491. doi:10.1101/cshperspect.a017491Pijnappel, W. W. M. P. (2001). The S. cerevisiae SET3 complex includes two histone deacetylases, Hos2 and Hst1, and is a meiotic-specific repressor of the sporulation gene program. Genes & Development, 15(22), 2991-3004. doi:10.1101/gad.207401Xie, J., Pierce, M., Gailus-Durner, V., Wagner, M., Winter, E., & Vershon, A. K. (1999). Sum1 and Hst1 repress middle sporulation-specific gene expression during mitosis in Saccharomyces cerevisiae. The EMBO Journal, 18(22), 6448-6454. doi:10.1093/emboj/18.22.6448Chalkiadaki, A., & Guarente, L. (2015). The multifaceted functions of sirtuins in cancer. Nature Reviews Cancer, 15(10), 608-624. doi:10.1038/nrc3985Roth, M., & Chen, W. Y. (2013). Sorting out functions of sirtuins in cancer. Oncogene, 33(13), 1609-1620. doi:10.1038/onc.2013.120Dolz-Edo, L., Rienzo, A., Poveda-Huertes, D., Pascual-Ahuir, A., & Proft, M. (2013). Deciphering Dynamic Dose Responses of Natural Promoters and Single cis Elements upon Osmotic and Oxidative Stress in Yeast. Molecular and Cellular Biology, 33(11), 2228-2240. doi:10.1128/mcb.00240-13Rienzo, A., Pascual-Ahuir, A., & Proft, M. (2012). The use of a real-time luciferase assay to quantify gene expression dynamics in the living yeast cell. Yeast, 29(6), 219-231. doi:10.1002/yea.290
Toxicity Mechanisms of the Food Contaminant Citrinin: Application of a Quantitative Yeast Model
Mycotoxins are important food contaminants and a serious threat for human nutrition. However, in many cases the mechanisms of toxicity for this diverse group of metabolites are poorly understood. Here we apply live cell gene expression reporters in yeast as a quantitative model to unravel the cellular defense mechanisms in response to the mycotoxin citrinin. We find that citrinin triggers a fast and dose dependent activation of stress responsive promoters such as GRE2 or SOD2. More specifically, oxidative stress responsive pathways via the transcription factors Yap1 and Skn7 are critically implied in the response to citrinin. Additionally, genes in various multidrug resistance transport systems are functionally involved in the resistance to citrinin. Our study identifies the antioxidant defense as a major physiological response in the case of citrinin. In general, our results show that the use of live cell gene expression reporters in yeast are a powerful tool to identify toxicity targets and detoxification mechanisms of a broad range of food contaminants relevant for human nutrition.This work was supported by Ministerio de Economia y Competitividad grant BFU2011-23326. We thank the Fond for Open Access Publication from Consejo Superior de Investigaciones Cientificas for supporting publication costs of this article.Pascual-Ahuir Giner, MD.; Vanacloig Pedrós, ME.; Proft, MH. (2014). Toxicity Mechanisms of the Food Contaminant Citrinin: Application of a Quantitative Yeast Model. Nutrients. 6(5):2077-2087. https://doi.org/10.3390/nu6052077S2077208765Moretti, A., Susca, A., Mulé, G., Logrieco, A. F., & Proctor, R. H. (2013). Molecular biodiversity of mycotoxigenic fungi that threaten food safety. International Journal of Food Microbiology, 167(1), 57-66. doi:10.1016/j.ijfoodmicro.2013.06.033Wu, F., Groopman, J. D., & Pestka, J. J. (2014). Public Health Impacts of Foodborne Mycotoxins. Annual Review of Food Science and Technology, 5(1), 351-372. doi:10.1146/annurev-food-030713-092431Bennett, J. W., & Klich, M. (2003). Mycotoxins. Clinical Microbiology Reviews, 16(3), 497-516. doi:10.1128/cmr.16.3.497-516.2003Flajs, D., & Peraica, M. (2009). Toxicological Properties of Citrinin. Archives of Industrial Hygiene and Toxicology, 60(4). doi:10.2478/10004-1254-60-2009-1992Bouslimi, A., Ouannes, Z., Golli, E. E., Bouaziz, C., Hassen, W., & Bacha, H. (2008). Cytotoxicity and Oxidative Damage in Kidney Cells Exposed to the Mycotoxins Ochratoxin A and Citrinin: Individual and Combined Effects. Toxicology Mechanisms and Methods, 18(4), 341-349. doi:10.1080/15376510701556682El Golli, E., Hassen, W., Bouslimi, A., Bouaziz, C., Ladjimi, M. M., & Bacha, H. (2006). Induction of Hsp 70 in Vero cells in response to mycotoxins. Toxicology Letters, 166(2), 122-130. doi:10.1016/j.toxlet.2006.06.004Kumar, R., Dwivedi, P. D., Dhawan, A., Das, M., & Ansari, K. M. (2011). Citrinin-Generated Reactive Oxygen Species Cause Cell Cycle Arrest Leading to Apoptosis via the Intrinsic Mitochondrial Pathway in Mouse Skin. Toxicological Sciences, 122(2), 557-566. doi:10.1093/toxsci/kfr143Ribeiro, S. M. R., Chagas, G. M., Campello, A. P., & Kluppel, M. L. W. (1997). Mechanism of citrinin-induced dysfunction of mitochondria. V. Effect on the homeostasis of the reactive oxygen species. Cell Biochemistry and Function, 15(3), 203-209. doi:10.1002/(sici)1099-0844(199709)15:33.0.co;2-jChen, C.-C., & Chan, W.-H. (2009). Inhibition of Citrinin-Induced Apoptotic Biochemical Signaling in Human Hepatoma G2 Cells by Resveratrol. International Journal of Molecular Sciences, 10(8), 3338-3357. doi:10.3390/ijms10083338Hsu, L.-C., Hsu, Y.-W., Liang, Y.-H., Lin, Z.-H., Kuo, Y.-H., & Pan, T.-M. (2012). Protective Effect of Deferricoprogen Isolated from Monascus purpureus NTU 568 on Citrinin-Induced Apoptosis in HEK-293 Cells. Journal of Agricultural and Food Chemistry, 60(32), 7880-7885. doi:10.1021/jf301889qIwahashi, H., Kitagawa, E., Suzuki, Y., Ueda, Y., Ishizawa, Y., Nobumasa, H., … Iwahashi, Y. (2007). Evaluation of toxicity of the mycotoxin citrinin using yeast ORF DNA microarray and Oligo DNA microarray. BMC Genomics, 8(1), 95. doi:10.1186/1471-2164-8-95(2012). Scientific Opinion on the risks for public and animal health related to the presence of citrinin in food and feed. EFSA Journal, 10(3). doi:10.2903/j.efsa.2012.2605Dos Santos, S. C. (2012). Yeast toxicogenomics: genome-wide responses to chemical stresses with impact in environmental health, pharmacology, and biotechnology. Frontiers in Genetics, 3. doi:10.3389/fgene.2012.00063Rienzo, A., Pascual-Ahuir, A., & Proft, M. (2012). The use of a real-time luciferase assay to quantify gene expression dynamics in the living yeast cell. Yeast, 29(6), 219-231. doi:10.1002/yea.2905Dolz-Edo, L., Rienzo, A., Poveda-Huertes, D., Pascual-Ahuir, A., & Proft, M. (2013). Deciphering Dynamic Dose Responses of Natural Promoters and Single cis Elements upon Osmotic and Oxidative Stress in Yeast. Molecular and Cellular Biology, 33(11), 2228-2240. doi:10.1128/mcb.00240-13Garay-Arroyo, A., & Covarrubias, A. A. (1999). Three genes whose expression is induced by stress inSaccharomyces cerevisiae. Yeast, 15(10A), 879-892. doi:10.1002/(sici)1097-0061(199907)15:10a3.0.co;2-qProft, M. (2001). Regulation of the Sko1 transcriptional repressor by the Hog1 MAP kinase in response to osmotic stress. The EMBO Journal, 20(5), 1123-1133. doi:10.1093/emboj/20.5.1123Mamnun, Y. M., Pandjaitan, R., Mahé, Y., Delahodde, A., & Kuchler, K. (2002). The yeast zinc finger regulators Pdr1p and Pdr3p control pleiotropic drug resistance (PDR) as homo- and heterodimers in vivo. Molecular Microbiology, 46(5), 1429-1440. doi:10.1046/j.1365-2958.2002.03262.xChagas, G. M., Campello, A. P., & Klüppel, M. L. W. (1992). Mechanism of citrinin-induced dysfunction of mitochondria. I. Effects on respiration, enzyme activities and membrane potential of renal cortical mitochondria. Journal of Applied Toxicology, 12(2), 123-129. doi:10.1002/jat.2550120209Klarić, M. Š., Želježić, D., Rumora, L., Peraica, M., Pepeljnjak, S., & Domijan, A.-M. (2011). A potential role of calcium in apoptosis and aberrant chromatin forms in porcine kidney PK15 cells induced by individual and combined ochratoxin A and citrinin. Archives of Toxicology, 86(1), 97-107. doi:10.1007/s00204-011-0735-9Yu, F.-Y., Liao, Y.-C., Chang, C.-H., & Liu, B.-H. (2006). Citrinin induces apoptosis in HL-60 cells via activation of the mitochondrial pathway. Toxicology Letters, 161(2), 143-151. doi:10.1016/j.toxlet.2005.08.009Dönmez-Altuntas, H., Dumlupinar, G., Imamoglu, N., Hamurcu, Z., & Liman, B. C. (2007). Effects of the mycotoxin citrinin on micronucleus formation in a cytokinesis-block genotoxicity assay in cultured human lymphocytes. Journal of Applied Toxicology, 27(4), 337-341. doi:10.1002/jat.1209Föllmann, W., Behm, C., & Degen, G. H. (2014). Toxicity of the mycotoxin citrinin and its metabolite dihydrocitrinone and of mixtures of citrinin and ochratoxin A in vitro. Archives of Toxicology, 88(5), 1097-1107. doi:10.1007/s00204-014-1216-8Knasmuller, S., Cavin, C., Chakraborty, A., Darroudi, F., Majer, B. J., Huber, W. W., & Ehrlich, V. A. (2004). Structurally Related Mycotoxins Ochratoxin A, Ochratoxin B, and Citrinin Differ in Their Genotoxic Activities and in Their Mode of Action in Human-Derived Liver (HepG2) Cells: Implications for Risk Assessment. Nutrition and Cancer, 50(2), 190-197. doi:10.1207/s15327914nc5002_9Thust, R., & Kneist, S. (1979). Activity of citrinin metabolized by rat and human microsome fractions in clastogenicity and SCE assays on Chinese hamster V79-E cells. Mutation Research/Genetic Toxicology, 67(4), 321-330. doi:10.1016/0165-1218(79)90028-4Van Loon, A. P., Pesold-Hurt, B., & Schatz, G. (1986). A yeast mutant lacking mitochondrial manganese-superoxide dismutase is hypersensitive to oxygen. Proceedings of the National Academy of Sciences, 83(11), 3820-3824. doi:10.1073/pnas.83.11.3820Delaunay, A., Isnard, A.-D., & Toledano, M. B. (2000). H2O2 sensing through oxidation of the Yap1 transcription factor. The EMBO Journal, 19(19), 5157-5166. doi:10.1093/emboj/19.19.5157Toone, W. M., Morgan, B. A., & Jones, N. (2001). Redox control of AP-1-like factors in yeast and beyond. Oncogene, 20(19), 2336-2346. doi:10.1038/sj.onc.1204384Heider, E. M., Harper, J. K., Grant, D. M., Hoffman, A., Dugan, F., Tomer, D. P., & O’Neill, K. L. (2006). Exploring unusual antioxidant activity in a benzoic acid derivative: a proposed mechanism for citrinin. Tetrahedron, 62(6), 1199-1208. doi:10.1016/j.tet.2005.10.06
Multi-layered control of peroxisomal activity upon salt stress in Saccharomyces cerevisiae
[EN] Peroxisomes are dynamic organelles and the sole location for fatty acid Ăź-oxidation in yeast cells. Here, we report that peroxisomal function is crucial for the adaptation to salt stress, especially upon sugar limitation. Upon stress, multiple layers of control regulate the activity and the number of peroxisomes. Activated Hog1 MAP kinase triggers the induction of genes encoding enzymes for fatty acid activation, peroxisomal import and Ăź-oxidation through the Adr1 transcriptional activator, which transiently associates with genes encoding fatty acid metabolic enzymes in a stress- and Hog1-dependent manner. Moreover, Na+ and Li+ stress increases the number of peroxisomes per cell in a Hog1-independent manner, which depends instead of the retrograde pathway and the dynamin related GTPases Dnm1 and Vps1. The strong activation of the Faa1 fatty acyl-CoA synthetase, which specifically localizes to lipid particles and peroxisomes, indicates that adaptation to salt stress requires the enhanced mobilization of fatty acids from internal lipid stores. Furthermore, the activation of mitochondrial respiration during stress depends on peroxisomes, mitochondrial acetyl-carnitine uptake is essential for salt resistance and the number of peroxisomes attached to the mitochondrial network increases during salt adaptation, which altogether indicates that stress-induced peroxisomal Ăź-oxidation triggers enhanced respiration upon salt shock.
peroxisomes, mitochondrial acetyl-carnitine uptake is essential for salt resistance, and the percentage of peroxisomes attached to the mitochondrial network increases during salt adaptation, which altogether indicates that stress-induced peroxisomal β-oxidation triggers enhanced respiration upon salt shock.We thank E. Hettema for the kind gift of GFP-PTS1 expressing yeast strains and E. de Nadal and F. Posas for the kind gift of Hog1-GFP wt and kinase dead alleles. This work was supported only in the initial phase by a grant from Ministerio de Economia y Competitividad (BFU2011-23326). The authors declare no conflict of interest. MP and AP-A conceived and designed the study, SM-E, JE-B, BA, MP and AP-A acquired, analyzed and interpreted data, MP wrote the manuscript.Manzanares-Estreder, S.; EspĂ Bardisa, J.; Pascual-Ahuir Giner, MD.; Proft, MH. (2016). Multi-layered control of peroxisomal activity upon salt stress in Saccharomyces cerevisiae. Molecular Microbiology. 104(5):851-868. https://doi.org/10.1111/mmi.13669S851868104
Nutraceuticals: opening the debate for a regulatory framework
Currently, nutraceuticals do not have a specific definition distinct from those of other food-derived categories, such as food supplements, herbal products, pre- and probiotics, functional foods, and fortified foods. Many studies have led to an understanding of the potential mechanisms of action of pharmaceutically active components contained in food that may improve health and reduce the risk of pathological conditions while enhancing overall well-being. Nevertheless, there is a lack of clear information and, often, the claimed health benefits may not be properly substantiated by safety and efficacy information or in vitro and in vivo data, which can induce false expectations and miss the target for a product to be effective, as claimed. An officially shared and accepted definition of nutraceuticals is still missing, as nutraceuticals are mostly referred to as pharma-foods, a powerful toolbox to be used beyond the diet but before the drugs to prevent and treat pathological conditions, such as in subjects who may not yet be eligible for conventional pharmaceutical therapy. Hence, it is of utmost importance to have a proper and unequivocal definition of nutraceuticals and shared regulations. It also seems wise to assess the safety, mechanism of action and efficacy of nutraceuticals with clinical data. A growing demand exists for nutraceuticals, which seem to reside in the grey area between pharmaceuticals and food. Nonetheless, given specific legislation from different countries, nutraceuticals are experiencing challenges with safety and health claim substantiation
Coordinated gene regulation in the initial phase of salt stress adaptation
This research was originally published in Journal of Biological Chemistry, 2015 - 16 : 10175- 10163 © the American Society for Biochemistry and Molecular Biology[EN] Stress triggers complex transcriptional responses, which include both gene activation and repression. We used time-resolved reporter assays in living yeast cells to gain insights into the coordination of positive and negative control of gene expression upon salt stress. We found that the repression of housekeeping genes coincides with the transient activation of defense genes and that the timing of this expression pattern depends on the severity of the stress. Moreover, we identified mutants that caused an alteration in the kinetics of this transcriptional control. Loss of function of the vacuolar H+-ATPase (vma1) or a defect in the biosynthesis of the osmolyte glycerol (gpd1) caused a prolonged repression of housekeeping genes and a delay in gene activation at inducible loci. Both mutants have a defect in the relocation of RNA polymerase II complexes at stress defense genes. Accordingly salt-activated transcription is delayed and less efficient upon partially respiratory growth conditions in which glycerol production is significantly reduced. Furthermore, the loss of Hog1 MAP kinase function aggravates the loss of RNA polymerase II from housekeeping loci, which apparently do not accumulate at inducible genes. Additionally the Def1 RNA polymerase II degradation factor, but not a high pool of nuclear polymerase II complexes, is needed for efficient stress-induced gene activation. The data presented here indicate that the finely tuned transcriptional control upon salt stress is dependent on physiological functions of the cell, such as the intracellular ion balance, the protective accumulation of osmolyte molecules, and the RNA polymerase II turnover.This work was supported by Ministerio de Economia y Competitividad Grant BFU2011-23326 (to M. P.).Vanacloig Pedros, ME.; Bets Plasencia, C.; Pascual-Ahuir Giner, MD.; Proft, MH. (2015). Coordinated gene regulation in the initial phase of salt stress adaptation. Journal of Biological Chemistry. 16(290):10163-10175. https://doi.org/10.1074/jbc.M115.637264S101631017516290De Nadal, E., Ammerer, G., & Posas, F. (2011). Controlling gene expression in response to stress. Nature Reviews Genetics, 12(12), 833-845. doi:10.1038/nrg3055Gasch, A. P., Spellman, P. T., Kao, C. M., Carmel-Harel, O., Eisen, M. B., Storz, G., … Brown, P. O. (2000). Genomic Expression Programs in the Response of Yeast Cells to Environmental Changes. Molecular Biology of the Cell, 11(12), 4241-4257. doi:10.1091/mbc.11.12.4241Gasch, A. P., & Werner-Washburne, M. (2002). The genomics of yeast responses to environmental stress and starvation. Functional & Integrative Genomics, 2(4-5), 181-192. doi:10.1007/s10142-002-0058-2Saito, H., & Posas, F. (2012). Response to Hyperosmotic Stress. Genetics, 192(2), 289-318. doi:10.1534/genetics.112.140863De Nadal, E., & Posas, F. (2009). Multilayered control of gene expression by stress-activated protein kinases. The EMBO Journal, 29(1), 4-13. doi:10.1038/emboj.2009.346Hohmann, S. (2009). Control of high osmolarity signalling in the yeast Saccharomyces cerevisiae. FEBS Letters, 583(24), 4025-4029. doi:10.1016/j.febslet.2009.10.069Nadal, E. d., Casadome, L., & Posas, F. (2003). Targeting the MEF2-Like Transcription Factor Smp1 by the Stress-Activated Hog1 Mitogen-Activated Protein Kinase. Molecular and Cellular Biology, 23(1), 229-237. doi:10.1128/mcb.23.1.229-237.2003MartĂnez-Montañés, F., Pascual-Ahuir, A., & Proft, M. (2010). Toward a Genomic View of the Gene Expression Program Regulated by Osmostress in Yeast. OMICS: A Journal of Integrative Biology, 14(6), 619-627. doi:10.1089/omi.2010.0046Proft, M. (2001). Regulation of the Sko1 transcriptional repressor by the Hog1 MAP kinase in response to osmotic stress. The EMBO Journal, 20(5), 1123-1133. doi:10.1093/emboj/20.5.1123Proft, M., & Serrano, R. (1999). Repressors and Upstream Repressing Sequences of the Stress-RegulatedENA1Gene inSaccharomyces cerevisiae: bZIP Protein Sko1p Confers HOG-Dependent Osmotic Regulation. Molecular and Cellular Biology, 19(1), 537-546. doi:10.1128/mcb.19.1.537Rep, M., Reiser, V., Gartner, U., Thevelein, J. M., Hohmann, S., Ammerer, G., & Ruis, H. (1999). Osmotic Stress-Induced Gene Expression inSaccharomyces cerevisiaeRequires Msn1p and the Novel Nuclear Factor Hot1p. Molecular and Cellular Biology, 19(8), 5474-5485. doi:10.1128/mcb.19.8.5474Ruiz-Roig, C., Noriega, N., Duch, A., Posas, F., & de Nadal, E. (2012). The Hog1 SAPK controls the Rtg1/Rtg3 transcriptional complex activity by multiple regulatory mechanisms. Molecular Biology of the Cell, 23(21), 4286-4296. doi:10.1091/mbc.e12-04-0289Westfall, P. J., Patterson, J. C., Chen, R. E., & Thorner, J. (2008). Stress resistance and signal fidelity independent of nuclear MAPK function. Proceedings of the National Academy of Sciences, 105(34), 12212-12217. doi:10.1073/pnas.0805797105Ariño, J., Aydar, E., Drulhe, S., Ganser, D., JorrĂn, J., Kahm, M., … Sychrová, H. (2014). Systems Biology of Monovalent Cation Homeostasis in Yeast. Advances in Microbial Systems Biology, 1-63. doi:10.1016/b978-0-12-800143-1.00001-4Hohmann, S. (2002). Osmotic adaptation in yeast-control of the yeast osmolyte system. Molecular Mechanisms of Water Transport Across Biological Membranes, 149-187. doi:10.1016/s0074-7696(02)15008-xHohmann, S., Krantz, M., & Nordlander, B. (2007). Yeast Osmoregulation. Osmosensing and Osmosignaling, 29-45. doi:10.1016/s0076-6879(07)28002-4Albertyn, J., Hohmann, S., Thevelein, J. M., & Prior, B. A. (1994). GPD1, which encodes glycerol-3-phosphate dehydrogenase, is essential for growth under osmotic stress in Saccharomyces cerevisiae, and its expression is regulated by the high-osmolarity glycerol response pathway. Molecular and Cellular Biology, 14(6), 4135-4144. doi:10.1128/mcb.14.6.4135Ansell, R., Granath, K., Hohmann, S., Thevelein, J. M., & Adler, L. (1997). The two isoenzymes for yeast NAD+-dependent glycerol 3-phosphate dehydrogenase encoded byGPD1andGPD2have distinct roles in osmoadaptation and redox regulation. The EMBO Journal, 16(9), 2179-2187. doi:10.1093/emboj/16.9.2179Norbeck, J., PĂĄhlman, A.-K., Akhtar, N., Blomberg, A., & Adler, L. (1996). Purification and Characterization of Two Isoenzymes of DL-Glycerol-3-phosphatase fromSaccharomyces cerevisiae. Journal of Biological Chemistry, 271(23), 13875-13881. doi:10.1074/jbc.271.23.13875Klipp, E., Nordlander, B., KrĂĽger, R., Gennemark, P., & Hohmann, S. (2005). Integrative model of the response of yeast to osmotic shock. Nature Biotechnology, 23(8), 975-982. doi:10.1038/nbt1114Proft, M., & Struhl, K. (2004). MAP Kinase-Mediated Stress Relief that Precedes and Regulates the Timing of Transcriptional Induction. Cell, 118(3), 351-361. doi:10.1016/j.cell.2004.07.016Li, S. C., Diakov, T. T., Rizzo, J. M., & Kane, P. M. (2011). Vacuolar H
+
-ATPase Works in Parallel with the HOG Pathway To Adapt Saccharomyces cerevisiae Cells to Osmotic Stress. Eukaryotic Cell, 11(3), 282-291. doi:10.1128/ec.05198-11Babazadeh, R., Adiels, C. B., Smedh, M., Petelenz-Kurdziel, E., Goksör, M., & Hohmann, S. (2013). Osmostress-Induced Cell Volume Loss Delays Yeast Hog1 Signaling by Limiting Diffusion Processes and by Hog1-Specific Effects. PLoS ONE, 8(11), e80901. doi:10.1371/journal.pone.0080901Miermont, A., Waharte, F., Hu, S., McClean, M. N., Bottani, S., Leon, S., & Hersen, P. (2013). Severe osmotic compression triggers a slowdown of intracellular signaling, which can be explained by molecular crowding. Proceedings of the National Academy of Sciences, 110(14), 5725-5730. doi:10.1073/pnas.1215367110Van Wuytswinkel, O., Reiser, V., Siderius, M., Kelders, M. C., Ammerer, G., Ruis, H., & Mager, W. H. (2000). Response of Saccharomyces cerevisiae to severe osmotic stress: evidence for a novel activation mechanism of the HOG MAP kinase pathway. Molecular Microbiology, 37(2), 382-397. doi:10.1046/j.1365-2958.2000.02002.xCook, K. E., & O’Shea, E. K. (2012). Hog1 Controls Global Reallocation of RNA Pol II upon Osmotic Shock in Saccharomyces cerevisiae. G3: Genes|Genomes|Genetics, 2(9), 1129-1136. doi:10.1534/g3.112.003251Nadal-Ribelles, M., Conde, N., Flores, O., González-Vallinas, J., Eyras, E., Orozco, M., … Posas, F. (2012). Hog1 bypasses stress-mediated down-regulation of transcription by RNA polymerase II redistribution and chromatin remodeling. Genome Biology, 13(11), R106. doi:10.1186/gb-2012-13-11-r106Dolz-Edo, L., Rienzo, A., Poveda-Huertes, D., Pascual-Ahuir, A., & Proft, M. (2013). Deciphering Dynamic Dose Responses of Natural Promoters and Single cis Elements upon Osmotic and Oxidative Stress in Yeast. Molecular and Cellular Biology, 33(11), 2228-2240. doi:10.1128/mcb.00240-13Berry, D. B., Guan, Q., Hose, J., Haroon, S., Gebbia, M., Heisler, L. E., … Gasch, A. P. (2011). Multiple Means to the Same End: The Genetic Basis of Acquired Stress Resistance in Yeast. PLoS Genetics, 7(11), e1002353. doi:10.1371/journal.pgen.1002353Guan, Q., Haroon, S., Bravo, D. G., Will, J. L., & Gasch, A. P. (2012). Cellular Memory of Acquired Stress Resistance inSaccharomyces cerevisiae. Genetics, 192(2), 495-505. doi:10.1534/genetics.112.143016Winzeler, E. A. (1999). Functional Characterization of the S. cerevisiae Genome by Gene Deletion and Parallel Analysis. Science, 285(5429), 901-906. doi:10.1126/science.285.5429.901Rienzo, A., Pascual-Ahuir, A., & Proft, M. (2012). The use of a real-time luciferase assay to quantify gene expression dynamics in the living yeast cell. Yeast, 29(6), 219-231. doi:10.1002/yea.2905Alberti, S., Gitler, A. D., & Lindquist, S. (2007). A suite of Gateway®cloning vectors for high-throughput genetic analysis inSaccharomyces cerevisiae. Yeast, 24(10), 913-919. doi:10.1002/yea.1502Aparicio, O., Geisberg, J. V., Sekinger, E., Yang, A., Moqtaderi, Z., & Struhl, K. (2005). Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific Genomic Sequences In Vivo. Current Protocols in Molecular Biology. doi:10.1002/0471142727.mb2103s69Woudstra, E. C., Gilbert, C., Fellows, J., Jansen, L., Brouwer, J., Erdjument-Bromage, H., … Svejstrup, J. Q. (2002). A Rad26–Def1 complex coordinates repair and RNA pol II proteolysis in response to DNA damage. Nature, 415(6874), 929-933. doi:10.1038/415929aCzeko, E., Seizl, M., Augsberger, C., Mielke, T., & Cramer, P. (2011). Iwr1 Directs RNA Polymerase II Nuclear Import. Molecular Cell, 42(2), 261-266. doi:10.1016/j.molcel.2011.02.033Esberg, A., Moqtaderi, Z., Fan, X., Lu, J., Struhl, K., & Byström, A. (2011). Iwr1 Protein Is Important for Preinitiation Complex Formation by All Three Nuclear RNA Polymerases in Saccharomyces cerevisiae. PLoS ONE, 6(6), e20829. doi:10.1371/journal.pone.0020829Rep, M., Albertyn, J., Thevelein, J. M., Prior, B. A., & Hohmann, S. (1999). Different signalling pathways contribute to the control of GPD1 gene expression by osmotic stress in Saccharomyces cerevisiae. Microbiology, 145(3), 715-727. doi:10.1099/13500872-145-3-715Bouwman, J., Kiewiet, J., Lindenbergh, A., van Eunen, K., Siderius, M., & Bakker, B. M. (2010). Metabolic regulation rather than de novo enzyme synthesis dominates the osmo-adaptation of yeast. Yeast, 28(1), 43-53. doi:10.1002/yea.1819Petelenz-Kurdziel, E., Kuehn, C., Nordlander, B., Klein, D., Hong, K.-K., Jacobson, T., … Klipp, E. (2013). Quantitative Analysis of Glycerol Accumulation, Glycolysis and Growth under Hyper Osmotic Stress. PLoS Computational Biology, 9(6), e1003084. doi:10.1371/journal.pcbi.1003084Dihazi, H., Kessler, R., & Eschrich, K. (2004). High Osmolarity Glycerol (HOG) Pathway-induced Phosphorylation and Activation of 6-Phosphofructo-2-kinase Are Essential for Glycerol Accumulation and Yeast Cell Proliferation under Hyperosmotic Stress. Journal of Biological Chemistry, 279(23), 23961-23968. doi:10.1074/jbc.m312974200Tamas, M. J., Luyten, K., Sutherland, F. C. W., Hernandez, A., Albertyn, J., Valadi, H., … Hohmann, S. (1999). Fps1p controls the accumulation and release of the compatible solute glycerol in yeast osmoregulation. Molecular Microbiology, 31(4), 1087-1104. doi:10.1046/j.1365-2958.1999.01248.xOstergaard, S., Olsson, L., Johnston, M., & Nielsen, J. (2000). Increasing galactose consumption by Saccharomyces cerevisiae through metabolic engineering of the GAL gene regulatory network. Nature Biotechnology, 18(12), 1283-1286. doi:10.1038/82400RIOS, G., FERRANDO, A., & SERRANO, R. (1997). Mechanisms of Salt Tolerance Conferred by Overexpression of theHAL1 Gene inSaccharomyces cerevisiae. Yeast, 13(6), 515-528. doi:10.1002/(sici)1097-0061(199705)13:63.0.co;2-xLi, S. C., & Kane, P. M. (2009). The yeast lysosome-like vacuole: Endpoint and crossroads. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1793(4), 650-663. doi:10.1016/j.bbamcr.2008.08.003Hamilton, C. A., Taylor, G. J., & Good, A. G. (2002). Vacuolar H+-ATPase, but not mitochondrial F1F0-ATPase, is required for NaCl tolerance inSaccharomyces cerevisiae. FEMS Microbiology Letters, 208(2), 227-232. doi:10.1111/j.1574-6968.2002.tb11086.xKellermayer, R. (2003). Extracellular Ca2+ sensing contributes to excess Ca2+ accumulation and vacuolar fragmentation in a pmr1Delta mutant of S. cerevisiae. Journal of Cell Science, 116(8), 1637-1646. doi:10.1242/jcs.00372Wilson, M. D., Harreman, M., Taschner, M., Reid, J., Walker, J., Erdjument-Bromage, H., … Svejstrup, J. Q. (2013). Proteasome-Mediated Processing of Def1, a Critical Step in the Cellular Response to Transcription Stress. Cell, 154(5), 983-995. doi:10.1016/j.cell.2013.07.028Somesh, B. P., Reid, J., Liu, W.-F., Søgaard, T. M. M., Erdjument-Bromage, H., Tempst, P., & Svejstrup, J. Q. (2005). Multiple Mechanisms Confining RNA Polymerase II Ubiquitylation to Polymerases Undergoing Transcriptional Arrest. Cell, 121(6), 913-923. doi:10.1016/j.cell.2005.04.01
Ask yeast how to burn your fats: lessons learned from the metabolic adaptation to salt stress
[EN] Here, we review and update the recent advances in the metabolic control during the adaptive response of budding yeast to hyperosmotic and salt stress, which is one of the best understood signaling events at the molecular level. This environmental stress can be easily applied and hence has been exploited in the past to generate an impressively detailed and comprehensive model of cellular adaptation. It is clear now that this stress modulates a great number of different physiological functions of the cell, which altogether contribute to cellular survival and adaptation. Primary defense mechanisms are the massive induction of stress tolerance genes in the nucleus, the activation of cation transport at the plasma membrane, or the production and intracellular accumulation of osmolytes. At the same time and in a coordinated manner, the cell shuts down the expression of housekeeping genes, delays the progression of the cell cycle, inhibits genomic replication, and modulates translation efficiency to optimize the response and to avoid cellular damage. To this fascinating interplay of cellular functions directly regulated by the stress, we have to add yet another layer of control, which is physiologically relevant for stress tolerance. Salt stress induces an immediate metabolic readjustment, which includes the up-regulation of peroxisomal biomass and activity in a coordinated manner with the reinforcement of mitochondrial respiratory metabolism. Our recent findings are consistent with a model, where salt stress triggers a metabolic shift from fermentation to respiration fueled by the enhanced peroxisomal oxidation of fatty acids. We discuss here the regulatory details of this stress-induced metabolic shift and its possible roles in the context of the previously known adaptive functions.The work of the authors was supported by
grants from Ministerio de EconomĂa y Competitividad (BFU2011-
23326 and BFU2016-75792-R).Pascual-Ahuir Giner, MD.; Manzanares-Estreder, S.; Timón Gómez, A.; Proft ., MH. (2017). Ask yeast how to burn your fats: lessons learned from the metabolic adaptation to salt stress. Current Genetics. 64(1):63-69. https://doi.org/10.1007/s00294-017-0724-5S6369641Aguilera J, Prieto JA (2001) The Saccharomyces cerevisiae aldose reductase is implied in the metabolism of methylglyoxal in response to stress conditions. Curr Genet 39:273–283Albertyn J, Hohmann S, Thevelein JM, Prior BA (1994) GPD1, which encodes glycerol-3-phosphate dehydrogenase, is essential for growth under osmotic stress in Saccharomyces cerevisiae, and its expression is regulated by the high-osmolarity glycerol response pathway. Mol Cell Biol 14:4135–4144Alepuz PM, Jovanovic A, Reiser V, Ammerer G (2001) Stress-induced map kinase Hog1 is part of transcription activation complexes. Mol Cell 7:767–777Alepuz PM, de Nadal E, Zapater M, Ammerer G, Posas F (2003) Osmostress-induced transcription by Hot1 depends on a Hog1-mediated recruitment of the RNA Pol II. EMBO J 22:2433–2442Ansell R, Granath K, Hohmann S, Thevelein JM, Adler L (1997) The two isoenzymes for yeast NAD+-dependent glycerol 3-phosphate dehydrogenase encoded by GPD1 and GPD2 have distinct roles in osmoadaptation and redox regulation. EMBO J 16:2179–2187Babazadeh R, Lahtvee PJ, Adiels CB, Goksor M, Nielsen JB, Hohmann S (2017) The yeast osmostress response is carbon source dependent. Sci Rep 7:990Bender T, Pena G, Martinou JC (2015) Regulation of mitochondrial pyruvate uptake by alternative pyruvate carrier complexes. EMBO J 34:911–924Berry DB, Gasch AP (2008) Stress-activated genomic expression changes serve a preparative role for impending stress in yeast. Mol Biol Cell 19:4580–4587Bilsland-Marchesan E, Arino J, Saito H, Sunnerhagen P, Posas F (2000) Rck2 kinase is a substrate for the osmotic stress-activated mitogen-activated protein kinase Hog1. Mol Cell Biol 20:3887–3895Brewster JL, Gustin MC (2014) Hog 1: 20 years of discovery and impact. Sci Signal 7:re7Clotet J, Posas F (2007) Control of cell cycle in response to osmostress: lessons from yeast. Methods Enzymol 428:63–76Clotet J, Escote X, Adrover MA, Yaakov G, Gari E, Aldea M, de Nadal E, Posas F (2006) Phosphorylation of Hsl1 by Hog1 leads to a G2 arrest essential for cell survival at high osmolarity. EMBO J 25:2338–2346Cook KE, O’Shea EK (2012) Hog1 controls global reallocation of RNA Pol II upon osmotic shock in Saccharomyces cerevisiae. Genes Genomes Genetics 2:1129–1136de Nadal E, Posas F (2015) Osmostress-induced gene expression—a model to understand how stress-activated protein kinases (SAPKs) regulate transcription. FEBS J 282:3275–3285de Nadal E, Alepuz PM, Posas F (2002) Dealing with osmostress through MAP kinase activation. EMBO Rep 3:735–740de Nadal E, Casadome L, Posas F (2003) Targeting the MEF2-like transcription factor Smp1 by the stress-activated Hog1 mitogen-activated protein kinase. Mol Cell Biol 23:229–237de Nadal E, Zapater M, Alepuz PM, Sumoy L, Mas G, Posas F (2004) The MAPK Hog1 recruits Rpd3 histone deacetylase to activate osmoresponsive genes. Nature 427:370–374Duch A, de Nadal E, Posas F (2013a) Dealing with transcriptional outbursts during S phase to protect genomic integrity. J Mol Biol 425:4745–4755Duch A, Felipe-Abrio I, Barroso S, Yaakov G, Garcia-Rubio M, Aguilera A, de Nadal E, Posas F (2013b) Coordinated control of replication and transcription by a SAPK protects genomic integrity. Nature 493:116–119Escote X, Zapater M, Clotet J, Posas F (2004) Hog1 mediates cell-cycle arrest in G1 phase by the dual targeting of Sic1. Nat Cell Biol 6:997–1002Ferreira C, van Voorst F, Martins A, Neves L, Oliveira R, Kielland-Brandt MC, Lucas C, Brandt A (2005) A member of the sugar transporter family, Stl1p is the glycerol/H+ symporter in Saccharomyces cerevisiae. Mol Biol Cell 16:2068–2076Gonzalez R, Morales P, Tronchoni J, Cordero-Bueso G, Vaudano E, Quiros M, Novo M, Torres-Perez R, Valero E (2016) New genes involved in osmotic stress tolerance in Saccharomyces cerevisiae. Front Microbiol 7:1545Ho YH, Gasch AP (2015) Exploiting the yeast stress-activated signaling network to inform on stress biology and disease signaling. Curr Genet 61:503–511Hohmann S (2015) An integrated view on a eukaryotic osmoregulation system. Curr Genet 61:373–382Hohmann S, Krantz M, Nordlander B (2007) Yeast osmoregulation. Methods Enzymol 428:29–45Hong SP, Carlson M (2007) Regulation of snf1 protein kinase in response to environmental stress. J Biol Chem 282:16838–16845Li SC, Diakov TT, Rizzo JM, Kane PM (2012) Vacuolar H+-ATPase works in parallel with the HOG pathway to adapt Saccharomyces cerevisiae cells to osmotic stress. Eukaryot Cell 11:282–291Maeta K, Izawa S, Inoue Y (2005) Methylglyoxal, a metabolite derived from glycolysis, functions as a signal initiator of the high osmolarity glycerol-mitogen-activated protein kinase cascade and calcineurin/Crz1-mediated pathway in Saccharomyces cerevisiae. J Biol Chem 280:253–260Manzanares-Estreder S, Espi-Bardisa J, Alarcon B, Pascual-Ahuir A, Proft M (2017) Multilayered control of peroxisomal activity upon salt stress in Saccharomyces cerevisiae. Mol Microbiol 104:851–868Mao K, Wang K, Zhao M, Xu T, Klionsky DJ (2011) Two MAPK-signaling pathways are required for mitophagy in Saccharomyces cerevisiae. J Cell Biol 193:755–767Martinez-Montanes F, Pascual-Ahuir A, Proft M (2010) Toward a genomic view of the gene expression program regulated by osmostress in yeast. OMICS 14:619–627Martinez-Pastor M, Proft M, Pascual-Ahuir A (2010) Adaptive changes of the yeast mitochondrial proteome in response to salt stress. OMICS 14:541–552Mas G, de Nadal E, Dechant R, Rodriguez de la Concepcion ML, Logie C, Jimeno-Gonzalez S, Chavez S, Ammerer G, Posas F (2009) Recruitment of a chromatin remodelling complex by the Hog1 MAP kinase to stress genes. EMBO J 28:326–336Mettetal JT, Muzzey D, Gomez-Uribe C, van Oudenaarden A (2008) The frequency dependence of osmo-adaptation in Saccharomyces cerevisiae. Science 319:482–484Molin C, Jauhiainen A, Warringer J, Nerman O, Sunnerhagen P (2009) mRNA stability changes precede changes in steady-state mRNA amounts during hyperosmotic stress. RNA 15:600–614Nadal-Ribelles M, Conde N, Flores O, Gonzalez-Vallinas J, Eyras E, Orozco M, de Nadal E, Posas F (2012) Hog1 bypasses stress-mediated down-regulation of transcription by RNA polymerase II redistribution and chromatin remodeling. Genome Biol 13:R106Pastor MM, Proft M, Pascual-Ahuir A (2009) Mitochondrial function is an inducible determinant of osmotic stress adaptation in yeast. J Biol Chem 284:30307–30317Petelenz-Kurdziel E, Kuehn C, Nordlander B, Klein D, Hong KK, Jacobson T, Dahl P, Schaber J, Nielsen J, Hohmann S, Klipp E (2013) Quantitative analysis of glycerol accumulation, glycolysis and growth under hyper osmotic stress. PLoS Comput Biol 9:e1003084Posas F, Chambers JR, Heyman JA, Hoeffler JP, de Nadal E, Arino J (2000) The transcriptional response of yeast to saline stress. J Biol Chem 275:17249–17255Proft M, Struhl K (2002) Hog1 kinase converts the Sko1-Cyc8-Tup1 repressor complex into an activator that recruits SAGA and SWI/SNF in response to osmotic stress. Mol Cell 9:1307–1317Proft M, Struhl K (2004) MAP kinase-mediated stress relief that precedes and regulates the timing of transcriptional induction. Cell 118:351–361Proft M, Pascual-Ahuir A, de Nadal E, Arino J, Serrano R, Posas F (2001) Regulation of the Sko1 transcriptional repressor by the Hog1 MAP kinase in response to osmotic stress. EMBO J 20:1123–1133Proft M, Mas G, de Nadal E, Vendrell A, Noriega N, Struhl K, Posas F (2006) The stress-activated Hog1 kinase is a selective transcriptional elongation factor for genes responding to osmotic stress. Mol Cell 23:241–250Ratnakumar S, Young ET (2010) Snf1 dependence of peroxisomal gene expression is mediated by Adr1. J Biol Chem 285:10703–10714Regot S, de Nadal E, Rodriguez-Navarro S, Gonzalez-Novo A, Perez-Fernandez J, Gadal O, Seisenbacher G, Ammerer G, Posas F (2013) The Hog1 stress-activated protein kinase targets nucleoporins to control mRNA export upon stress. J Biol Chem 288:17384–17398Rep M, Krantz M, Thevelein JM, Hohmann S (2000) The transcriptional response of Saccharomyces cerevisiae to osmotic shock. Hot1p and Msn2p/Msn4p are required for the induction of subsets of high osmolarity glycerol pathway-dependent genes. J Biol Chem 275:8290–8300Rep M, Proft M, Remize F, Tamas M, Serrano R, Thevelein JM, Hohmann S (2001) The Saccharomyces cerevisiae Sko1p transcription factor mediates HOG pathway-dependent osmotic regulation of a set of genes encoding enzymes implicated in protection from oxidative damage. Mol Microbiol 40:1067–1083Rienzo A, Poveda-Huertes D, Aydin S, Buchler NE, Pascual-Ahuir A, Proft M (2015) Different mechanisms confer gradual control and memory at nutrient- and stress-regulated genes in yeast. Mol Cell Biol 35:3669–3683Romero-Santacreu L, Moreno J, Perez-Ortin JE, Alepuz P (2009) Specific and global regulation of mRNA stability during osmotic stress in Saccharomyces cerevisiae. RNA 15:1110–1120Roy A, Hashmi S, Li Z, Dement AD, Cho KH, Kim JH (2016) The glucose metabolite methylglyoxal inhibits expression of the glucose transporter genes by inactivating the cell surface glucose sensors Rgt2 and Snf3 in yeast. Mol Biol Cell 27:862–871Ruiz-Roig C, Noriega N, Duch A, Posas F, de Nadal E (2012) The Hog1 SAPK controls the Rtg1/Rtg3 transcriptional complex activity by multiple regulatory mechanisms. Mol Biol Cell 23:4286–4296Saito H, Posas F (2012) Response to hyperosmotic stress. Genetics 192:289–318Sekito T, Thornton J, Butow RA (2000) Mitochondria-to-nuclear signaling is regulated by the subcellular localization of the transcription factors Rtg1p and Rtg3p. Mol Biol Cell 11:2103–2115Silva RD, Sotoca R, Johansson B, Ludovico P, Sansonetty F, Silva MT, Peinado JM, Corte-Real M (2005) Hyperosmotic stress induces metacaspase- and mitochondria-dependent apoptosis in Saccharomyces cerevisiae. Mol Microbiol 58:824–834Sole C, Nadal-Ribelles M, de Nadal E, Posas F (2015) A novel role for lncRNAs in cell cycle control during stress adaptation. Curr Genet 61:299–308Tamas MJ, Luyten K, Sutherland FC, Hernandez A, Albertyn J, Valadi H, Li H, Prior BA, Kilian SG, Ramos J, Gustafsson L, Thevelein JM, Hohmann S (1999) Fps1p controls the accumulation and release of the compatible solute glycerol in yeast osmoregulation. Mol Microbiol 31:1087–1104Teige M, Scheikl E, Reiser V, Ruis H, Ammerer G (2001) Rck2, a member of the calmodulin-protein kinase family, links protein synthesis to high osmolarity MAP kinase signaling in budding yeast. Proc Natl Acad Sci USA 98:5625–5630Timon-Gomez A, Proft M, Pascual-Ahuir A (2013) Differential regulation of mitochondrial pyruvate carrier genes modulates respiratory capacity and stress tolerance in yeast. PLoS One 8:e79405Vanacloig-Pedros E, Bets-Plasencia C, Pascual-Ahuir A, Proft M (2015) Coordinated gene regulation in the initial phase of salt stress adaptation. J Biol Chem 290:10163–10175Warringer J, Hult M, Regot S, Posas F, Sunnerhagen P (2010) The HOG pathway dictates the short-term translational response after hyperosmotic shock. Mol Biol Cell 21:3080–3092Wei CJ, Tanner RD, Malaney GW (1982) Effect of sodium chloride on bakers’ yeast growing in gelatin. Appl Environ Microbiol 43:757–763Westfall PJ, Patterson JC, Chen RE, Thorner J (2008) Stress resistance and signal fidelity independent of nuclear MAPK function. Proc Natl Acad Sci USA 105:12212–12217Ye T, Garcia-Salcedo R, Ramos J, Hohmann S (2006) Gis4, a new component of the ion homeostasis system in the yeast Saccharomyces cerevisiae. Eukaryot Cell 5:1611–1621Yoshida A, Wei D, Nomura W, Izawa S, Inoue Y (2012) Reduction of glucose uptake through inhibition of hexose transporters and enhancement of their endocytosis by methylglyoxal in Saccharomyces cerevisiae. J Biol Chem 287:701–71
Live-cell assays reveal selectivity and sensitivity of the multidrug response in budding yeast
14 páginas, 9 figuras. Figuras S1 y S2, Tablas S1-S3 en material suplementario: http://www.jbc.org/content/suppl/2019/07/11/RA119.009291.DC1/152730_2_supp_360630_pqfm3f.pdfPleiotropic drug resistance arises by the enhanced extrusion of bioactive molecules and is present in a wide range of organisms, ranging from fungi to human cells. A key feature of this adaptation is the sensitive detection of intracellular xenobiotics by transcriptional activators, activating expression of multiple drug exporters. Here, we investigated the selectivity and sensitivity of the budding yeast (Saccharomyces cerevisiae) multidrug response to better understand how differential drug recognition leads to specific activation of drug exporter genes and to drug resistance. Applying live-cell luciferase reporters, we demonstrate that the SNQ2, PDR5, PDR15, and YOR1 transporter genes respond to different mycotoxins, menadione, and hydrogen peroxide in a distinguishable manner and with characteristic amplitudes, dynamics, and sensitivities. These responses correlated with differential sensitivities of the respective transporter mutants to the specific xenobiotics. We further establish a binary vector system, enabling quantitative determination of xenobiotic-transcription factor (TF) interactions in real time. Applying this system we found that the TFs Pdr1, Pdr3, Yrr1, Stb5, and Pdr8 have largely different drug recognition patterns. We noted that Pdr1 is the most promiscuous activator, whereas Yrr1 and Stb5 are selective for ochratoxin A and hydrogen peroxide, respectively. We also show that Pdr1 is rapidly degraded after xenobiotic exposure, which leads to a desensitization of the Pdr1-specific response upon repeated activation. The findings of our work indicate that in the yeast multidrug system, several transcriptional activators with distinguishable selectivities trigger differential activation of the transporter genes.This work was supported by Ministerio de EconomĂa y Competitividad Grant
BFU2016-75792-R. The authors declare that they have no conflicts of interest with the contents of this article.Peer reviewe