Interactions of netrin-1 through its glycosylation sites immobilize Deleted in Colorectal Cancer (DCC) by favoring its constitutive clustering

Netrin-1 is a protein that attracts neurons expressing the membrane receptor Deleted in Colorectal Cancer DCC. In colon carcinoma, the interaction between netrin-1 and DCC prevents apoptosis. Crystallographic data suggest that these processes involve the clustering of DCC, the observation of which in cells remains elusive, as do the molecular determinants of DCC-netrin-1 interactions and their impact on DCC organization and mobility. To address these questions, we used fluorescence photobleaching, single-particle tracking and super-resolution techniques to characterize DCC organization and mobility on the cell surface. Our results show that netrin-1 impedes DCC mobility in the plasma membrane by promoting the growth of constitutive DCC nanoclusters at the expense of free DCC. Furthermore, we show that these effects are mediated primarily by the three glycosylation sites in the LamVI domain of netrin-1 and, to a lesser extent, by the C-terminal domain and its RGD binding site.