Depletion of COPI in cancer cells: the role of reactive oxygen species in the induction of lipid accumulation, noncanonical lipophagy and apoptosis.

The coatomer protein complex 1 (COPI) is a multisubunit complex that coats intracellular vesicles and is involved in intracellular protein trafficking. Recently we and others found that depletion of COPI complex subunits zeta (COPZ1) and delta (ARCN1) preferentially kills tumor cells relative to normal cells. Here we delineate the specific cellular effects and sequence of events of COPI complex depletion in tumor cells. We find that this depletion leads to the inhibition of mitochondrial oxidative phosphorylation and the elevation of reactive oxygen species (ROS) production, followed by accumulation of lipid droplets (LDs) and autophagy-associated proteins LC3-II and SQSTM1/p62 and, finally, apoptosis of the tumor cells. Inactivation of ROS in COPI-depleted cells with the mitochondrial-specific quencher, mitoquinone mesylate, attenuated apoptosis and markedly decreased both the size and the number of LDs. COPI depletion caused ROS-dependent accumulation of LC3-II and SQSTM1 which colocalizes with LDs. Lack of double-membrane autophagosomes and insensitivity to Atg5 deletion suggested an accumulation of a microlipophagy complex on the surface of LDs induced by depletion of the COPI complex. Our findings suggest a sequence of cellular events triggered by COPI depletion, starting with inhibition of oxidative phosphorylation, followed by ROS activation and accumulation of LDs and apoptosis.

TßRIII/ß-arrestin2 regulates integrin α5ß1 trafficking, function, and localization in epithelial cells.

The type III TGF-ß receptor (TßRIII) is a ubiquitous co-receptor for TGF-ß superfamily ligands with roles in suppressing cancer progression, in part through suppressing cell motility. Here we demonstrate that TßRIII promotes epithelial cell adhesion to fibronectin in a ß-arrestin2 dependent and TGF-ß/BMP independent manner by complexing with active integrin α5ß1, and mediating ß-arrestin2-dependent α5ß1 internalization and trafficking to nascent focal adhesions. TßRIII-mediated integrin α5ß1 trafficking regulates cell adhesion and fibronectin fibrillogenesis in epithelial cells, as well as α5 localization in breast cancer patients. We further demonstrate that increased TßRIII expression correlates with increased α5 localization at sites of cell-cell adhesion in breast cancer patients, while higher TßRIII expression is a strong predictor of overall survival in breast cancer patients. These data support a novel, clinically relevant role for TßRIII in regulating integrin α5 localization, reveal a novel crosstalk mechanism between the integrin and TGF-ß superfamily signaling pathways and identify ß-arrestin2 as a regulator of α5ß1 trafficking.