Nuclear translocation of MLKL enhances necroptosis by a RIP1/RIP3-independent mechanism in H9c2 cardiomyoblasts
Accumulating evidence suggests that necroptosis of cardiomyocytes contributes to cardiovascular diseases. Lethal disruption of the plasma membrane in necroptosis is induced by oligomers of mixed lineage kinase domain-like (MLKL) that is translocated to the membrane from the cytosol. However, the rol...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-02-01
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| Series: | Journal of Pharmacological Sciences |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S1347861322000974 |
| _version_ | 1797943067770093568 |
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| author | Shoya Ino Toshiyuki Yano Atsushi Kuno Masaya Tanno Hidemichi Kouzu Tatsuya Sato Tomohisa Yamashita Wataru Ohwada Arata Osanami Toshifumi Ogawa Yuki Toda Masaki Shimizu Tetsuji Miura |
| author_facet | Shoya Ino Toshiyuki Yano Atsushi Kuno Masaya Tanno Hidemichi Kouzu Tatsuya Sato Tomohisa Yamashita Wataru Ohwada Arata Osanami Toshifumi Ogawa Yuki Toda Masaki Shimizu Tetsuji Miura |
| author_sort | Shoya Ino |
| collection | DOAJ |
| description | Accumulating evidence suggests that necroptosis of cardiomyocytes contributes to cardiovascular diseases. Lethal disruption of the plasma membrane in necroptosis is induced by oligomers of mixed lineage kinase domain-like (MLKL) that is translocated to the membrane from the cytosol. However, the role played by cytoplasmic-nuclear shuttling of MLKL is unclear. Here, we tested the hypothesis that translocation of MLKL to the nucleus promotes the necroptosis of cardiomyocytes. Activation of the canonical necroptotic signaling pathway by a combination of TNF-α and zVAD (TNF/zVAD) increased nuclear MLKL levels in a RIP1-activity-dependent manner in H9c2 cells, a rat cardiomyoblast cell line. By use of site-directed mutagenesis, we found a nuclear export signal sequence in MLKL and prepared its mutant (MLKL-L280/283/284A), though a search for a nuclear import signal was unsuccessful. MLKL-L280/283/284A localized to both the cytosol and the nucleus. Expression of MLKL-L280/283/284A induced necroptotic cell death, which was attenuated by GppNHp, an inhibitor of Ran-mediated nuclear import, but not by inhibition of RIP1 activity or knockdown of RIP3 expression. GppNHp partly suppressed H9c2 cell death induced by TNF/zVAD treatment. These results suggest that MLKL that is translocated to the nucleus via RIP1-mediated necroptotic signaling enhances the necroptosis of cardiomyocytes through a RIP1-/RIP3-independent mechanism. |
| first_indexed | 2024-04-10T20:17:30Z |
| format | Article |
| id | doaj.art-281af19d393a476a9faf3bcbc6aff414 |
| institution | Directory Open Access Journal |
| issn | 1347-8613 |
| language | English |
| last_indexed | 2024-04-10T20:17:30Z |
| publishDate | 2023-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Pharmacological Sciences |
| spelling | doaj.art-281af19d393a476a9faf3bcbc6aff4142023-01-26T04:44:42ZengElsevierJournal of Pharmacological Sciences1347-86132023-02-011512134143Nuclear translocation of MLKL enhances necroptosis by a RIP1/RIP3-independent mechanism in H9c2 cardiomyoblastsShoya Ino0Toshiyuki Yano1Atsushi Kuno2Masaya Tanno3Hidemichi Kouzu4Tatsuya Sato5Tomohisa Yamashita6Wataru Ohwada7Arata Osanami8Toshifumi Ogawa9Yuki Toda10Masaki Shimizu11Tetsuji Miura12Department of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan; Department of Pharmacology, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan; Department of Cell Physiology and Signal Transduction, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, JapanDepartment of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan; Department of Clinical Pharmacology, Faculty of Pharmaceutical Sciences, Hokkaido University of Science, Sapporo, Japan; Corresponding author. Department of Cardiovascular, Renal, and Metabolic Medicine Sapporo Medical University School of Medicine South-1, West-16, Chuo-ku, Sapporo 060-8543, Japan.Accumulating evidence suggests that necroptosis of cardiomyocytes contributes to cardiovascular diseases. Lethal disruption of the plasma membrane in necroptosis is induced by oligomers of mixed lineage kinase domain-like (MLKL) that is translocated to the membrane from the cytosol. However, the role played by cytoplasmic-nuclear shuttling of MLKL is unclear. Here, we tested the hypothesis that translocation of MLKL to the nucleus promotes the necroptosis of cardiomyocytes. Activation of the canonical necroptotic signaling pathway by a combination of TNF-α and zVAD (TNF/zVAD) increased nuclear MLKL levels in a RIP1-activity-dependent manner in H9c2 cells, a rat cardiomyoblast cell line. By use of site-directed mutagenesis, we found a nuclear export signal sequence in MLKL and prepared its mutant (MLKL-L280/283/284A), though a search for a nuclear import signal was unsuccessful. MLKL-L280/283/284A localized to both the cytosol and the nucleus. Expression of MLKL-L280/283/284A induced necroptotic cell death, which was attenuated by GppNHp, an inhibitor of Ran-mediated nuclear import, but not by inhibition of RIP1 activity or knockdown of RIP3 expression. GppNHp partly suppressed H9c2 cell death induced by TNF/zVAD treatment. These results suggest that MLKL that is translocated to the nucleus via RIP1-mediated necroptotic signaling enhances the necroptosis of cardiomyocytes through a RIP1-/RIP3-independent mechanism.http://www.sciencedirect.com/science/article/pii/S1347861322000974NecroptosisCardiomyocyteMLKLRIP1RIP3 |
| spellingShingle | Shoya Ino Toshiyuki Yano Atsushi Kuno Masaya Tanno Hidemichi Kouzu Tatsuya Sato Tomohisa Yamashita Wataru Ohwada Arata Osanami Toshifumi Ogawa Yuki Toda Masaki Shimizu Tetsuji Miura Nuclear translocation of MLKL enhances necroptosis by a RIP1/RIP3-independent mechanism in H9c2 cardiomyoblasts Journal of Pharmacological Sciences Necroptosis Cardiomyocyte MLKL RIP1 RIP3 |
| title | Nuclear translocation of MLKL enhances necroptosis by a RIP1/RIP3-independent mechanism in H9c2 cardiomyoblasts |
| title_full | Nuclear translocation of MLKL enhances necroptosis by a RIP1/RIP3-independent mechanism in H9c2 cardiomyoblasts |
| title_fullStr | Nuclear translocation of MLKL enhances necroptosis by a RIP1/RIP3-independent mechanism in H9c2 cardiomyoblasts |
| title_full_unstemmed | Nuclear translocation of MLKL enhances necroptosis by a RIP1/RIP3-independent mechanism in H9c2 cardiomyoblasts |
| title_short | Nuclear translocation of MLKL enhances necroptosis by a RIP1/RIP3-independent mechanism in H9c2 cardiomyoblasts |
| title_sort | nuclear translocation of mlkl enhances necroptosis by a rip1 rip3 independent mechanism in h9c2 cardiomyoblasts |
| topic | Necroptosis Cardiomyocyte MLKL RIP1 RIP3 |
| url | http://www.sciencedirect.com/science/article/pii/S1347861322000974 |
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