Crumbs controls epithelial integrity by inhibiting Rac1 and PI3K

Abstract

Drosophila Crumbs (Crb) and its mammalian ortholog CRB3 control epithelial polarity through poorly understood molecular mechanisms. Elucidating these mechanisms is crucial, because the physiology of epithelia largely depends on the polarized architecture of individual epithelial cells. In addition, loss of CRB3 favors tumor cell growth, metastasis and epithelial to mesenchymal transition (EMT). Using Drosophila embryos, we report that Rac1 sustains PI3K signaling, which is required for Rac1 activation. Crb represses this positive-feedback loop. Notably, this property confers to Crb its ability to promote epithelial integrity in vivo, because attenuation of either Rac1 or PI3K activity rescues the crb mutant phenotype. Moreover, inhibition of Rac1 or PI3K results in Crb-dependent apical membrane growth, whereas Rac1 activation restricts membrane localization of Crb and interferes with apical domain formation. This illustrates that Crb and the Rac1-PI3K module are antagonists, and that the fine balance between the activities of these proteins is crucial to maintain epithelial organization and an appropriate apical to basolateral ratio. Together, our results elucidate a mechanism that mediates Crb function and further define the role of PI3K and Rac1 in epithelial morphogenesis, allowing for a better understanding of how distinct membrane domains are regulated in polarized epithelial cells.