Vinculin Y822 is an important determinant of ligand binding

Abstract

Vinculin is an actin-binding protein present at cell–matrix and cell–cell adhesions, which plays a critical role in bearing force experienced by cells and dissipating it onto the cytoskeleton. Recently, we identified a key tyrosine residue, Y822, whose phosphorylation plays a critical role in force transmission at cell–cell adhesions. The role of Y822 in human cancer remains unknown, even though Y822 is mutated to Y822C in uterine cancers. Here, we investigated the effect of this amino acid substitution and that of a phosphodeficient Y822F vinculin in cancer cells. We observed that the presence of the Y822C mutation led to cells that proliferate and migrate more rapidly and contained smaller focal adhesions when compared to cells with wild-type vinculin. In contrast, the presence of the Y822F mutation led to highly spread cells with larger focal adhesions and increased contractility. Furthermore, we provide evidence that Y822C vinculin forms a disulfide bond with paxillin, accounting for some of the elevated phosphorylated paxillin recruitment. Taken together, these data suggest that vinculin Y822 modulates the recruitment of ligands.

Vinculin-Arp2/3 Interaction Inhibits Branched Actin Assembly to Control Cell Migration and Cell Cycle Progression

“…In 2D wound healing assay of MCF10A cells, vinculin KO cells were more efficient at closing the wound than parental cells, as previously reported using 4T1 cells (DeWane et al , 2022). KO MCF10A cells exhibited fast and directional migration towards the wound and transmitted the signal further back in the monolayer, indicating that the mechanotransduction of E-Cadherin dependent cell adhesions that vinculin provides (le Duc et al , 2010) is not essential to this transmission, and even rather inhibitory.…”

“…Vinculin belongs to both cell adhesions, the specific incorporation into cell-cell adhesions being determined by Abl-mediated phosphorylation of Y822 (Bays et al, 2014). Decreased cell-cell adhesion upon vinculin KO was recently observed in mouse 4T1 breast cancer line (DeWane et al, 2022) and is in line with the aberrant AJs between cardiac myocytes reported in heart-specific KO of vinculin in mice (Zemljic-Harpf et al, 2007). Vinculin is an essential component of cell-cell junctions that allows myosin-dependent tensile forces to develop mature junctions (Twiss et al, 2012).…”

“…Force-dependent recruitment of vinculin results in its activation allowing vinculin tail to link additional actin filaments and thereby reinforce the cytoskeletal attachment of cell adhesions. In the process, vinculin mechanotransduces signals, since vinculin-depleted cells exhibit enhanced proliferation, survival and anchorage-independent growth (Fernández et al , 1993; Subauste et al , 2004; DeWane et al , 2022).…”

“…The copyright holder for this preprint this version posted October 9, 2023. ; https://doi.org/10.1101/2023.10.09.561480 doi: bioRxiv preprint vinculin-depleted cells exhibit enhanced proliferation, survival and anchorage-independent growth (Fernández et al, 1993;Subauste et al, 2004;DeWane et al, 2022).…”