Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamics
The intermediate filament protein vimentin constitutes a critical sensor for electrophilic and oxidative stress, which induce extensive reorganization of the vimentin cytoskeletal network. Here, we have investigated the mechanisms underlying these effects. In vitro, electrophilic lipids, including 1...
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| Format: | Article |
| Language: | English |
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Elsevier
2019-05-01
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| Series: | Redox Biology |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231718310127 |
| _version_ | 1818924280116674560 |
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| author | Andreia Mónico Sofia Duarte María A. Pajares Dolores Pérez-Sala |
| author_facet | Andreia Mónico Sofia Duarte María A. Pajares Dolores Pérez-Sala |
| author_sort | Andreia Mónico |
| collection | DOAJ |
| description | The intermediate filament protein vimentin constitutes a critical sensor for electrophilic and oxidative stress, which induce extensive reorganization of the vimentin cytoskeletal network. Here, we have investigated the mechanisms underlying these effects. In vitro, electrophilic lipids, including 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) and 4-hydroxynonenal (HNE), directly bind to vimentin, whereas the oxidant diamide induces disulfide bond formation. Mutation of the single vimentin cysteine residue (Cys328) blunts disulfide formation and reduces lipoxidation by 15d-PGJ2, but not HNE. Preincubation with these agents differentially hinders NaCl-induced filament formation by wild-type vimentin, with effects ranging from delayed elongation and increased filament diameter to severe impairment of assembly or aggregation. Conversely, the morphology of vimentin Cys328Ser filaments is mildly or not affected. Interestingly, preformed vimentin filaments are more resistant to electrophile-induced disruption, although chemical modification is not diminished, showing that vimentin (lip)oxidation prior to assembly is more deleterious. In cells, electrophiles, particularly diamide, induce a fast and drastic disruption of existing filaments, which requires the presence of Cys328. As the cellular vimentin network is under continuous remodeling, we hypothesized that vimentin exchange on filaments would be necessary for diamide-induced disruption. We confirmed that strategies reducing vimentin dynamics, as monitored by FRAP, including cysteine crosslinking and ATP synthesis inhibition, prevent diamide effect. In turn, phosphorylation may promote vimentin disassembly. Indeed, treatment with the phosphatase inhibitor calyculin A to prevent dephosphorylation intensifies electrophile-induced wild-type vimentin filament disruption. However, whereas a phosphorylation-deficient vimentin mutant is only partially protected from disorganization, Cys328Ser vimentin is virtually resistant, even in the presence of calyculin A. Together, these results indicate that modification of Cys328 and vimentin exchange are critical for electrophile-induced network disruption. Keywords: Vimentin, Lipoxidation, HNE, Cyclopentenone prostaglandins, Intermediate filaments, Vimentin filament morphology and dynamics, Vimentin oxidation |
| first_indexed | 2024-12-20T02:22:49Z |
| format | Article |
| id | doaj.art-f91a731fe8a54b3981235cf407597795 |
| institution | Directory Open Access Journal |
| issn | 2213-2317 |
| language | English |
| last_indexed | 2024-12-20T02:22:49Z |
| publishDate | 2019-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj.art-f91a731fe8a54b3981235cf4075977952022-12-21T19:56:47ZengElsevierRedox Biology2213-23172019-05-0123Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamicsAndreia Mónico0Sofia Duarte1María A. Pajares2Dolores Pérez-Sala3Department of Structural and Chemical Biology, Centro de Investigaciones Biológicas (CSIC), Ramiro de Maeztu 9, 28040 Madrid, SpainDepartment of Structural and Chemical Biology, Centro de Investigaciones Biológicas (CSIC), Ramiro de Maeztu 9, 28040 Madrid, SpainDepartment of Structural and Chemical Biology, Centro de Investigaciones Biológicas (CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain; Molecular Hepatology Group, Instituto de Investigación Sanitaria del Hospital Universitario La Paz (IdiPAZ), Paseo de la Castellana 261, 28046 Madrid, SpainDepartment of Structural and Chemical Biology, Centro de Investigaciones Biológicas (CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain; Correspondence to: Department of Structural and Chemical Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas (C.S.I.C.), Ramiro de Maeztu, 9, 28040 Madrid, Spain.The intermediate filament protein vimentin constitutes a critical sensor for electrophilic and oxidative stress, which induce extensive reorganization of the vimentin cytoskeletal network. Here, we have investigated the mechanisms underlying these effects. In vitro, electrophilic lipids, including 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) and 4-hydroxynonenal (HNE), directly bind to vimentin, whereas the oxidant diamide induces disulfide bond formation. Mutation of the single vimentin cysteine residue (Cys328) blunts disulfide formation and reduces lipoxidation by 15d-PGJ2, but not HNE. Preincubation with these agents differentially hinders NaCl-induced filament formation by wild-type vimentin, with effects ranging from delayed elongation and increased filament diameter to severe impairment of assembly or aggregation. Conversely, the morphology of vimentin Cys328Ser filaments is mildly or not affected. Interestingly, preformed vimentin filaments are more resistant to electrophile-induced disruption, although chemical modification is not diminished, showing that vimentin (lip)oxidation prior to assembly is more deleterious. In cells, electrophiles, particularly diamide, induce a fast and drastic disruption of existing filaments, which requires the presence of Cys328. As the cellular vimentin network is under continuous remodeling, we hypothesized that vimentin exchange on filaments would be necessary for diamide-induced disruption. We confirmed that strategies reducing vimentin dynamics, as monitored by FRAP, including cysteine crosslinking and ATP synthesis inhibition, prevent diamide effect. In turn, phosphorylation may promote vimentin disassembly. Indeed, treatment with the phosphatase inhibitor calyculin A to prevent dephosphorylation intensifies electrophile-induced wild-type vimentin filament disruption. However, whereas a phosphorylation-deficient vimentin mutant is only partially protected from disorganization, Cys328Ser vimentin is virtually resistant, even in the presence of calyculin A. Together, these results indicate that modification of Cys328 and vimentin exchange are critical for electrophile-induced network disruption. Keywords: Vimentin, Lipoxidation, HNE, Cyclopentenone prostaglandins, Intermediate filaments, Vimentin filament morphology and dynamics, Vimentin oxidationhttp://www.sciencedirect.com/science/article/pii/S2213231718310127 |
| spellingShingle | Andreia Mónico Sofia Duarte María A. Pajares Dolores Pérez-Sala Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamics Redox Biology |
| title | Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamics |
| title_full | Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamics |
| title_fullStr | Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamics |
| title_full_unstemmed | Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamics |
| title_short | Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamics |
| title_sort | vimentin disruption by lipoxidation and electrophiles role of the cysteine residue and filament dynamics |
| url | http://www.sciencedirect.com/science/article/pii/S2213231718310127 |
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