{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T23:34:53Z","timestamp":1767828893637,"version":"3.49.0"},"reference-count":45,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,10,21]],"date-time":"2020-10-21T00:00:00Z","timestamp":1603238400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Regional Development Fund (ERDF) through the COMPETE e Operational Competitiveness Program and national funds through FCT (Foundation for Science and Technology) grant to E. Valerio (SFRH\/BPD\/ 75922\/2011).","award":["Strategic Funding UIDB\/04423\/2020 and UI : SFRH\/BPD\/ 75922\/2011; UIDB\/04423\/2020 and UIDP\/04423\/2020 ; No 823860"],"award-info":[{"award-number":["Strategic Funding UIDB\/04423\/2020 and UI : SFRH\/BPD\/ 75922\/2011; UIDB\/04423\/2020 and UIDP\/04423\/2020 ; No 823860"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Microcystins (MCs) are hepatotoxins produced by some cyanobacteria. They are cyclic peptides that inhibit the serine\/threonine protein phosphatases (PPs) PP1 and PP2A, especially PP2A. The inhibition of PP2A triggers a series of molecular events, which are responsible for most MC cytotoxic and genotoxic effects on animal cells. It is also known that MCs induce oxidative stress in cells due to the production of reactive oxygen species (ROS). However, a complete characterization of the toxic effects of MCs is still not accomplished. This study aimed to clarify additional molecular mechanisms involved in MC-LR toxicity, using Saccharomyces cerevisiae as eukaryotic model organism. First, a shotgun proteomic analysis of S. cerevisiae VL3 cells response to 1 nM, 10 nM, 100 nM, and 1 \u03bcM MC-LR was undertaken and compared to the control (cells not exposed to MC-LR). This analysis revealed a high number of proteins differentially expressed related with gene translation and DNA replication stress; oxidative stress; cell cycle regulation and carbohydrate metabolism. Inference of genotoxic effects of S. cerevisiae VL3 cells exposed to different concentrations of MC-LR were evaluated by analyzing the expression of genes Apn1, Apn2, Rad27, Ntg1, and Ntg2 (from the Base Excision Repair (BER) DNA repair system) using the Real-Time RT-qPCR technique. These genes displayed alterations after exposure to MC-LR, particularly the Apn1\/Apn2\/Rad27, pointing out effects of MC-LR in the Base Excision Repair system (BER). Overall, this study supports the role of oxidative stress and DNA replication stress as important molecular mechanisms of MC-LR toxicity. Moreover, this study showed that even at low-concentration, MC-LR can induce significant changes in the yeast proteome and in gene expression.<\/jats:p>","DOI":"10.3390\/toxins12100667","type":"journal-article","created":{"date-parts":[[2020,10,23]],"date-time":"2020-10-23T02:01:42Z","timestamp":1603418502000},"page":"667","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["New Insights in Saccharomyces cerevisiae Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene Expression"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8189-244X","authenticated-orcid":false,"given":"Elisabete","family":"Val\u00e9rio","sequence":"first","affiliation":[{"name":"Departamento de Sa\u00fade Ambiental, Instituto Nacional de Sa\u00fade Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016 Lisboa, Portugal"}]},{"given":"Sara","family":"Barreiros","sequence":"additional","affiliation":[{"name":"Departamento de Sa\u00fade Ambiental, Instituto Nacional de Sa\u00fade Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016 Lisboa, Portugal"}]},{"given":"Sara","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Departamento de Sa\u00fade Ambiental, Instituto Nacional de Sa\u00fade Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016 Lisboa, Portugal"}]},{"given":"Maria V.","family":"Turkina","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Clinical Sciences, Faculty of Medicine and Clinical Sciences, Link\u00f6ping University, 581 83 Link\u00f6ping, Sweden"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3585-2417","authenticated-orcid":false,"given":"Vitor M.","family":"Vasconcelos","sequence":"additional","affiliation":[{"name":"Centro Interdisciplinar de Investiga\u00e7\u00e3o Marinha e Ambiental (CIIMAR\/CIMAR), University of Porto, Terminal de Cruzeiros do Porto de Leix\u00f5es, Av. General Norton de Matos, s\/n, 4450-208 Matosinhos, Portugal"},{"name":"Biology Department, Faculty of Sciences, University of Porto, Rua do Campo Alegre, s\/n, 4169-007 Porto, Portugal"}]},{"given":"Alexandre","family":"Campos","sequence":"additional","affiliation":[{"name":"Centro Interdisciplinar de Investiga\u00e7\u00e3o Marinha e Ambiental (CIIMAR\/CIMAR), University of Porto, Terminal de Cruzeiros do Porto de Leix\u00f5es, Av. General Norton de Matos, s\/n, 4450-208 Matosinhos, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Meriluoto, J., Spoof, L., and Codd, G.A. (2017). Microcystins and Nodularins. Handbook of Cyanobacterial Monitoring and Cyanotoxin Analysis, John Wiley & Sons, Ltd.. [1st ed.].","DOI":"10.1002\/9781119068761"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1002\/mnfr.200600185","article-title":"Toxins of cyanobacteria","volume":"51","author":"Speijers","year":"2007","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"268","DOI":"10.3390\/ijms11010268","article-title":"Molecular Mechanisms of Microcystin Toxicity in Animal Cells","volume":"11","author":"Campos","year":"2010","journal-title":"Int. J. Mol. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1032","DOI":"10.2174\/1389557516666160219130553","article-title":"new insights on the mode of action of microcystins in animal cells\u2014A Review","volume":"16","author":"Vasconcelos","year":"2016","journal-title":"Mini Rev. Med. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1007\/s12403-019-00303-1","article-title":"A Comparative review of the effect of microcystin-LR on the proteome","volume":"12","author":"Welten","year":"2020","journal-title":"Expo Health"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1039\/C7TX00043J","article-title":"Toxic mechanisms of microcystins in mammals","volume":"6","author":"McLellan","year":"2017","journal-title":"Toxicol. Res. (Camb.)"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1016\/j.toxicon.2015.05.005","article-title":"Mechanisms of microcystin-LR-induced cytoskeletal disruption in animal cells","volume":"101","author":"Zhou","year":"2015","journal-title":"Toxicon"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Jiang, J., Shan, Z., Xu, W., Wang, X., Zhou, J., Kong, D., and Xu, J. (2013). Microcystin-LR induced reactive oxygen species mediate cytoskeletal disruption and apoptosis of hepatocytes in Cyprinus carpio L.. PloS ONE, 8.","DOI":"10.1371\/journal.pone.0084768"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1016\/j.toxicon.2009.10.003","article-title":"Differential oxidative stress responses to pure Microcystin-LR and Microcystin-containing and non-containing cyanobacterial crude extracts on Caco-2 cells","volume":"55","author":"Puerto","year":"2010","journal-title":"Toxicon"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Zong, W., Zhang, S., Wang, Q., Teng, Y., Liu, Y., and Du, Y. (2018). Evaluation of the direct and indirect regulation pathways of glutathione target to the hepatotoxicity of Microcystin-LR. BioMed. Res. Int.","DOI":"10.1155\/2018\/5672637"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0378-1097(03)00100-9","article-title":"Role of oxidative stress and mitochondrial changes in cyanobacteria-induced apoptosis and hepatotoxicity","volume":"220","author":"Ding","year":"2003","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"4722","DOI":"10.3390\/ijms12074722","article-title":"Microcystin-LR induces apoptosis via NF-\u03baB\/iNOS pathway in INS-1 cells","volume":"12","author":"Ji","year":"2011","journal-title":"Int. J. Mol. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1016\/j.jes.2014.07.036","article-title":"Role of nitric oxide in the genotoxic response to chronic microcystin-LR exposure in human\u2013hamster hybrid cells","volume":"29","author":"Wang","year":"2015","journal-title":"J. Environ. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"110096","DOI":"10.1016\/j.ecoenv.2019.110096","article-title":"Microcystins-LR induced apoptosis via S-nitrosylation of GAPDH in colorectal cancer cells","volume":"190","author":"Li","year":"2020","journal-title":"Ecotoxicol. Environ. Saf."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.toxicon.2014.08.059","article-title":"Effects of microcystin-LR on Saccharomyces cerevisiae growth, oxidative stress and apoptosis","volume":"90","author":"Vilares","year":"2014","journal-title":"Toxicon"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Gowder, S. (2013). The kidney Vero-E6 cell line: A suitable model to study the toxicity of microcystins. New Insights into Toxicity and Drug Testing, InTech.","DOI":"10.5772\/55886"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.toxicon.2016.01.059","article-title":"Proteomic and Real-Time PCR analyses of Saccharomyces cerevisiae VL3 exposed to microcystin-LR reveals a set of protein alterations transversal to several eukaryotic models","volume":"112","author":"Campos","year":"2016","journal-title":"Toxicon"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"109824","DOI":"10.1016\/j.ecoenv.2019.109824","article-title":"Responses of the proteome in testis of mice exposed chronically to environmentally relevant concentrations of Microcystin-LR","volume":"187","author":"Zhou","year":"2020","journal-title":"Ecotoxicol. Environ. Saf."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"113019","DOI":"10.1016\/j.envpol.2019.113019","article-title":"Proteomic analysis of the hepatotoxicity of Microcystis aeruginosa in adult zebrafish (Danio rerio) and its potential mechanisms","volume":"254","author":"Du","year":"2019","journal-title":"Environ. Pollut."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1007\/s00204-016-1681-3","article-title":"Prolonged exposure to low-dose microcystin induces nonalcoholic steatohepatitis in mice: A systems toxicology study","volume":"91","author":"He","year":"2017","journal-title":"Arch. Toxicol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Friedberg, E.C., Walker, G.C., Siede, W., Wood, R.D., Schultz, R.A., and Ellenberger, T. (2006). DNA Repair and Mutagenesis, ASM Press.","DOI":"10.1128\/9781555816704"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1534\/genetics.112.145219","article-title":"DNA repair mechanisms and the bypass of DNA damage in Saccharomyces cerevisiae","volume":"193","author":"Boiteux","year":"2013","journal-title":"Genetics"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1002\/(SICI)1522-7278(199902)14:1<61::AID-TOX10>3.0.CO;2-R","article-title":"Microcystin-LR and liver tumor promotion: Effects on cytokinesis, ploidy, and apoptosis in cultured hepatocytes","volume":"14","author":"Humpage","year":"1999","journal-title":"Environ. Toxicol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1689","DOI":"10.1016\/j.tiv.2010.05.018","article-title":"Microcystin-LR activates the ERK1\/2 kinases and stimulates the proliferation of the monkey kidney-derived cell line Vero-E6","volume":"24","author":"Dias","year":"2010","journal-title":"Toxicol. In Vitro"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/j.toxicon.2009.04.014","article-title":"Morphological and ultrastructural effects of microcystin-LR from Microcystis aeruginosa extract on a kidney cell line","volume":"54","author":"Alverca","year":"2010","journal-title":"Toxicon"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.tiv.2012.09.009","article-title":"Involvement of endoplasmic reticulum and autophagy in microcystin-LR toxicity in Vero-E6 and HepG2 cell lines","volume":"27","author":"Menezes","year":"2013","journal-title":"Toxicol. In Vitro"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"D607","DOI":"10.1093\/nar\/gky1131","article-title":"STRING v11: Protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets","volume":"47","author":"Szklarczyk","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/j.toxicon.2009.01.029","article-title":"Comparative study of the cytotoxic effect of microcistin-LR and purified extracts from Microcystis aeruginosa on a kidney cell line","volume":"53","author":"Dias","year":"2009","journal-title":"Toxicon"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/S0045-6535(99)00402-6","article-title":"Study on the cytotoxicity of microcystin-LR on cultured cells","volume":"41","author":"Chong","year":"2000","journal-title":"Chemosphere"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3582","DOI":"10.1091\/mbc.e12-05-0413","article-title":"The essential iron-sulfur protein Rli1 is an important target accounting for inhibition of cell growth by reactive oxygen species","volume":"23","author":"Alhebshi","year":"2012","journal-title":"Mol. Biol. Cell."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e20934","DOI":"10.7554\/eLife.20934","article-title":"Eukaryotic translation initiation factor 3 plays distinct roles at the mRNA entry and exit channels of the ribosomal preinitiation complex","volume":"5","author":"Aitken","year":"2016","journal-title":"eLife"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5981","DOI":"10.1093\/nar\/gkh934","article-title":"Roles of Rad23 protein in yeast nucleotide excision repair","volume":"32","author":"Xie","year":"2004","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2631","DOI":"10.1091\/mbc.11.8.2631","article-title":"Thioredoxin peroxidase is required for the transcriptional response to oxidative stress in budding yeast","volume":"11","author":"Ross","year":"2000","journal-title":"Mol. Biol. Cell."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1007\/s10541-005-0102-7","article-title":"Mitochondrial metabolism of reactive oxygen species","volume":"70","author":"Andreyev","year":"2005","journal-title":"Biochemistry (Mosc.)"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1016\/j.cels.2017.02.011","article-title":"Systematic analysis of human protein phosphatase interactions and dynamics","volume":"4","author":"Yadav","year":"2017","journal-title":"Cell Syst."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1046\/j.1474-9728.2003.00041.x","article-title":"Base excision repair activities required for yeast to attain a full chronological life span","volume":"2","author":"Maclean","year":"2003","journal-title":"Aging Cell"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/0076-6879(91)94004-V","article-title":"Getting started with yeast","volume":"194","author":"Sherman","year":"1991","journal-title":"Methods Enzymol."},{"key":"ref_38","unstructured":"Seidel, C. (2019, June 30). OD660 vs Number of Cells. Available online: http:\/\/www.pangloss.com\/seidel\/Protocols\/ODvsCells.html."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1038\/nmeth.1322","article-title":"Universal sample preparation method for proteome analysis","volume":"6","author":"Zougman","year":"2009","journal-title":"Nat. Methods"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.jprot.2016.01.010","article-title":"Shotgun proteomics to unravel marine mussel (Mytilus edulis) response to long-term exposure to low salinity and propranolol in a Baltic Sea microcosm","volume":"137","author":"Campos","year":"2016","journal-title":"J. Proteom."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"956","DOI":"10.1093\/nar\/27.4.956","article-title":"Relationships between yeast Rad27 and Apn1 in response to apurinic\/apyrimidinic (AP) sites in DNA","volume":"27","author":"Wu","year":"1999","journal-title":"Nucleic Acids Res."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Teste, M., Duquenne, M., Fran\u00e7ois, J.M., and Parrou, J. (2009). Validation of reference genes for quantitative expression analysis by real-time RT-PCR in Saccharomyces cerevisiae. BMC Mol. Biol., 10.","DOI":"10.1186\/1471-2199-10-99"},{"key":"ref_43","unstructured":"Frank, H. (2010). Chapter 9-The real-time polymerase chain reaction (RT-PCR). Calculations for Molecular Biology and Biotechnology, Academic Press. [2nd ed.]."},{"key":"ref_44","unstructured":"(2018, January 31). Multiple Experiment Viewer. Available online: https:\/\/www.mybiosoftware.com\/mev-4-6-2-multiple-experiment-viewer.html."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2002","DOI":"10.1093\/nar\/29.9.e45","article-title":"A new mathematical model for relative quantification in real-time RT\u2013PCR","volume":"29","author":"Pfaffl","year":"2001","journal-title":"Nucleic Acids Res."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/12\/10\/667\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:25:36Z","timestamp":1760178336000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/12\/10\/667"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,21]]},"references-count":45,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2020,10]]}},"alternative-id":["toxins12100667"],"URL":"https:\/\/doi.org\/10.3390\/toxins12100667","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,10,21]]}}}