New Insights in <i>Saccharomyces cerevisiae</i> Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene Expression
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 genot...
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MDPI AG
2020-10-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/12/10/667 |
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| author | Elisabete Valério Sara Barreiros Sara Rodrigues Maria V. Turkina Vitor M. Vasconcelos Alexandre Campos |
| author_facet | Elisabete Valério Sara Barreiros Sara Rodrigues Maria V. Turkina Vitor M. Vasconcelos Alexandre Campos |
| author_sort | Elisabete Valério |
| collection | DOAJ |
| description | 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 <i>Saccharomyces cerevisiae</i> as eukaryotic model organism. First, a shotgun proteomic analysis of <i>S. cerevisiae</i> VL3 cells response to 1 nM, 10 nM, 100 nM, and 1 μM 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 <i>S. cerevisiae</i> VL3 cells exposed to different concentrations of MC-LR were evaluated by analyzing the expression of genes <i>Apn1</i>, <i>Apn2</i>, <i>Rad27</i>, <i>Ntg1</i>, and <i>Ntg2</i> (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 <i>Apn1</i>/<i>Apn2</i>/<i>Rad27</i>, 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. |
| first_indexed | 2024-03-10T15:26:29Z |
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| id | doaj.art-4cec3444a1714e8cb4d1ec0c99ec48e7 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-10T15:26:29Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-4cec3444a1714e8cb4d1ec0c99ec48e72023-11-20T18:00:26ZengMDPI AGToxins2072-66512020-10-01121066710.3390/toxins12100667New Insights in <i>Saccharomyces cerevisiae</i> Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene ExpressionElisabete Valério0Sara Barreiros1Sara Rodrigues2Maria V. Turkina3Vitor M. Vasconcelos4Alexandre Campos5Departamento de Saúde Ambiental, Instituto Nacional de Saúde Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016 Lisboa, PortugalDepartamento de Saúde Ambiental, Instituto Nacional de Saúde Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016 Lisboa, PortugalDepartamento de Saúde Ambiental, Instituto Nacional de Saúde Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016 Lisboa, PortugalDepartment of Biomedical and Clinical Sciences, Faculty of Medicine and Clinical Sciences, Linköping University, 581 83 Linköping, SwedenCentro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR/CIMAR), University of Porto, Terminal de Cruzeiros do Porto de Leixões, Av. General Norton de Matos, s/n, 4450-208 Matosinhos, PortugalCentro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR/CIMAR), University of Porto, Terminal de Cruzeiros do Porto de Leixões, Av. General Norton de Matos, s/n, 4450-208 Matosinhos, PortugalMicrocystins (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 <i>Saccharomyces cerevisiae</i> as eukaryotic model organism. First, a shotgun proteomic analysis of <i>S. cerevisiae</i> VL3 cells response to 1 nM, 10 nM, 100 nM, and 1 μM 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 <i>S. cerevisiae</i> VL3 cells exposed to different concentrations of MC-LR were evaluated by analyzing the expression of genes <i>Apn1</i>, <i>Apn2</i>, <i>Rad27</i>, <i>Ntg1</i>, and <i>Ntg2</i> (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 <i>Apn1</i>/<i>Apn2</i>/<i>Rad27</i>, 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.https://www.mdpi.com/2072-6651/12/10/667Microcystin-LR<i>Saccharomyces cerevisiae</i>MTT assaysshotgun proteomicsBER genesReal-Time RT-qPCR |
| spellingShingle | Elisabete Valério Sara Barreiros Sara Rodrigues Maria V. Turkina Vitor M. Vasconcelos Alexandre Campos New Insights in <i>Saccharomyces cerevisiae</i> Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene Expression Toxins Microcystin-LR <i>Saccharomyces cerevisiae</i> MTT assays shotgun proteomics BER genes Real-Time RT-qPCR |
| title | New Insights in <i>Saccharomyces cerevisiae</i> Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene Expression |
| title_full | New Insights in <i>Saccharomyces cerevisiae</i> Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene Expression |
| title_fullStr | New Insights in <i>Saccharomyces cerevisiae</i> Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene Expression |
| title_full_unstemmed | New Insights in <i>Saccharomyces cerevisiae</i> Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene Expression |
| title_short | New Insights in <i>Saccharomyces cerevisiae</i> Response to the Cyanotoxin Microcystin-LR, Revealed by Proteomics and Gene Expression |
| title_sort | new insights in i saccharomyces cerevisiae i response to the cyanotoxin microcystin lr revealed by proteomics and gene expression |
| topic | Microcystin-LR <i>Saccharomyces cerevisiae</i> MTT assays shotgun proteomics BER genes Real-Time RT-qPCR |
| url | https://www.mdpi.com/2072-6651/12/10/667 |
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