Vimentin filament organization and stress sensing depend on its single cysteine residue and zinc binding

Abstract

The vimentin filament network plays a key role in cell architecture and signalling, as well as in epithelial–mesenchymal transition. Vimentin C328 is targeted by various oxidative modifications, but its role in vimentin organization is not known. Here we show that C328 is essential for vimentin network reorganization in response to oxidants and electrophiles, and is required for optimal vimentin performance in network expansion, lysosomal distribution and aggresome formation. C328 may fulfil these roles through interaction with zinc. In vitro, micromolar zinc protects vimentin from iodoacetamide modification and elicits vimentin polymerization into optically detectable structures; in cells, zinc closely associates with vimentin and its depletion causes reversible filament disassembly. Finally, zinc transport-deficient human fibroblasts show increased vimentin solubility and susceptibility to disruption, which are restored by zinc supplementation. These results unveil a critical role of C328 in vimentin organization and open new perspectives for the regulation of intermediate filaments by zinc.

A vimentin binding small molecule leads to mitotic disruption in mesenchymal cancers

“…Further, treating FOXC2-HMLER cells with FiVe1 dramatically reduced the appearance of dibromobimane-crosslinked dimeric VIM protein content by Western blotting, an assay which has previously been used to monitor the filamentous status of VIM in cells and in solution (SI Appendix, Fig. S5K) (24). Additionally, treatment of FOXC2-HMLER cells with FiVe1 or WIF-A induced the dosedependent accumulation of lower molecular weight VIM degradation products as visualized by Western blotting (Fig.…”

Senescent cells expose and secrete an oxidized form of membrane-bound vimentin as revealed by a natural polyreactive antibody

“…C328 in human vimentin has been shown to be the target of electrophilic lipids that include cyclopentenone prostaglandins (cyPG), which are reactive lipids that are generated under conditions of inflammation and oxidative stress (49,81,82). C328 is also subject to covalent addition of the reactive aldehyde HNE, which is similar to MDA (48). CML was previously shown to modify lysines in the exposed linker regions of vimentin after UV treatment (44), and although we detected CML-modified vimentin by IP-Western, our proteomic screen failed to identify the location of these modifications.…”

“…CML was previously shown to modify lysines in the exposed linker regions of vimentin after UV treatment (44), and although we detected CML-modified vimentin by IP-Western, our proteomic screen failed to identify the location of these modifications. Importantly, at the cellular level, the modification of vimentin by these oxidation-associated modifications (cyPG, HNE, and CML) has been shown to result in the disruption of the intermediate filament network and the generation of intracellular aggresomes (44,48,82).…”

“…S6D). Interestingly, C328 has been shown to be required for proper function of vimentin in human cells under normal and oxidative conditions (48). The oxidative adduct aldehyde 4-hydroxynonenal (HNE) has been previously shown to modify vimentin at C328 (49); however, an HNE modification at C328 was not identified in our proteomic screen.…”

Proteomic analysis of human glutathione transferase omega (hGSTO1) stable transfection in a 6-hydroxydopamine-induced neuronal cells

“…Glutathionylation of vimentin has been reported in several studies (Townsend et al 2003;Townsend 2007;Xiong et al 2011). Vimentin possesses only a single cysteine residue at position Cys 328 (Perez-Sala et al 2015). This residue has a role in filament formation, vimentin network extension and subunit exchange, organelle positioning etc.…”