Characterization of the RAS/RAF/ERK Signal Cascade as a Novel Regulating Factor in Alpha-Amanitin-Induced Cytotoxicity in Huh-7 Cells
The well-known hepatotoxicity mechanism resulting from alpha-amanitin (α-AMA) exposure arises from RNA polymerase II (RNAP II) inhibition. RNAP Ⅱ inhibition occurs through the dysregulation of mRNA synthesis. However, the signaling pathways in hepatocytes that arise from α-AMA have not yet been full...
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| Format: | Article |
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MDPI AG
2022-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/20/12294 |
| _version_ | 1797472773919997952 |
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| author | Doeun Kim Min Seo Lee Eunji Sung Sangkyu Lee Hye Suk Lee |
| author_facet | Doeun Kim Min Seo Lee Eunji Sung Sangkyu Lee Hye Suk Lee |
| author_sort | Doeun Kim |
| collection | DOAJ |
| description | The well-known hepatotoxicity mechanism resulting from alpha-amanitin (α-AMA) exposure arises from RNA polymerase II (RNAP II) inhibition. RNAP Ⅱ inhibition occurs through the dysregulation of mRNA synthesis. However, the signaling pathways in hepatocytes that arise from α-AMA have not yet been fully elucidated. Here, we identified that the RAS/RAF/ERK signaling pathway was activated through quantitative phosphoproteomic and molecular biological analyses in Huh-7 cells. Bioinformatics analysis showed that α-AMA exposure increased protein phosphorylation in a time-dependent α-AMA exposure. In addition, phosphorylation increased not only the components of the ERK signaling pathway but also U2AF65 and SPF45, known splicing factors. Therefore, we propose a novel mechanism of α-AMA as follows. The RAS/RAF/ERK signaling pathway involved in aberrant splicing events is activated by α-AMA exposure followed by aberrant splicing events leading to cell death in Huh-7 cells. |
| first_indexed | 2024-03-09T20:06:57Z |
| format | Article |
| id | doaj.art-a43fbb5441354655a4919cb545735978 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T20:06:57Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-a43fbb5441354655a4919cb5457359782023-11-24T00:30:35ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-10-0123201229410.3390/ijms232012294Characterization of the RAS/RAF/ERK Signal Cascade as a Novel Regulating Factor in Alpha-Amanitin-Induced Cytotoxicity in Huh-7 CellsDoeun Kim0Min Seo Lee1Eunji Sung2Sangkyu Lee3Hye Suk Lee4Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu 41566, KoreaBK21 FOUR-Sponsored Advanced Program for SmartPharma Leaders, College of Pharmacy, The Catholic University of Korea, Bucheon 14662, KoreaBK21 FOUR Community-Based Intelligent Novel Drug Discovery Education Unit, College of Pharmacy, Kyungpook National University, Daegu 41566, KoreaResearch Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu 41566, KoreaBK21 FOUR-Sponsored Advanced Program for SmartPharma Leaders, College of Pharmacy, The Catholic University of Korea, Bucheon 14662, KoreaThe well-known hepatotoxicity mechanism resulting from alpha-amanitin (α-AMA) exposure arises from RNA polymerase II (RNAP II) inhibition. RNAP Ⅱ inhibition occurs through the dysregulation of mRNA synthesis. However, the signaling pathways in hepatocytes that arise from α-AMA have not yet been fully elucidated. Here, we identified that the RAS/RAF/ERK signaling pathway was activated through quantitative phosphoproteomic and molecular biological analyses in Huh-7 cells. Bioinformatics analysis showed that α-AMA exposure increased protein phosphorylation in a time-dependent α-AMA exposure. In addition, phosphorylation increased not only the components of the ERK signaling pathway but also U2AF65 and SPF45, known splicing factors. Therefore, we propose a novel mechanism of α-AMA as follows. The RAS/RAF/ERK signaling pathway involved in aberrant splicing events is activated by α-AMA exposure followed by aberrant splicing events leading to cell death in Huh-7 cells.https://www.mdpi.com/1422-0067/23/20/12294alpha-amanitintoxic mushroomacute liver failureglobal phosphoproteomeRAS/RAF/ERK signaling pathway |
| spellingShingle | Doeun Kim Min Seo Lee Eunji Sung Sangkyu Lee Hye Suk Lee Characterization of the RAS/RAF/ERK Signal Cascade as a Novel Regulating Factor in Alpha-Amanitin-Induced Cytotoxicity in Huh-7 Cells International Journal of Molecular Sciences alpha-amanitin toxic mushroom acute liver failure global phosphoproteome RAS/RAF/ERK signaling pathway |
| title | Characterization of the RAS/RAF/ERK Signal Cascade as a Novel Regulating Factor in Alpha-Amanitin-Induced Cytotoxicity in Huh-7 Cells |
| title_full | Characterization of the RAS/RAF/ERK Signal Cascade as a Novel Regulating Factor in Alpha-Amanitin-Induced Cytotoxicity in Huh-7 Cells |
| title_fullStr | Characterization of the RAS/RAF/ERK Signal Cascade as a Novel Regulating Factor in Alpha-Amanitin-Induced Cytotoxicity in Huh-7 Cells |
| title_full_unstemmed | Characterization of the RAS/RAF/ERK Signal Cascade as a Novel Regulating Factor in Alpha-Amanitin-Induced Cytotoxicity in Huh-7 Cells |
| title_short | Characterization of the RAS/RAF/ERK Signal Cascade as a Novel Regulating Factor in Alpha-Amanitin-Induced Cytotoxicity in Huh-7 Cells |
| title_sort | characterization of the ras raf erk signal cascade as a novel regulating factor in alpha amanitin induced cytotoxicity in huh 7 cells |
| topic | alpha-amanitin toxic mushroom acute liver failure global phosphoproteome RAS/RAF/ERK signaling pathway |
| url | https://www.mdpi.com/1422-0067/23/20/12294 |
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