Fat-regulated adaptor protein Dlish binds the growth suppressor Expanded and controls its stability and ubiquitination

Significance

To regulate the growth and size of organs, cells can use information from their neighbors to modify intracellular mediators of cell proliferation. The intracellular Hippo pathway is a widely utilized nexus for growth control in animals, but its regulation by extracellular signals is not fully understood. We here identify a pathway that regulates organ size in Drosophila, triggered by the transmembrane receptor, the giant protocadherin Fat. We show that the Fat-regulated SH3 domain adaptor protein Dlish binds to and reduces the stability of the growth suppressor Expanded, a known regulator of the Hippo pathway. The destabilization of Expanded by Dlish works in parallel to a previously established pathway in which Dlish increases levels of the growth-stimulating protein Dachs.

Abstract

The Drosophila protocadherin Fat controls organ size through the Hippo pathway, but the biochemical links to the Hippo pathway components are still poorly defined. We previously identified Dlish, an SH3 domain protein that physically interacts with Fat and the type XX myosin Dachs, and showed that Fat’s regulation of Dlish levels and activity helps limit Dachs-mediated inhibition of Hippo pathway activity. We here characterize a parallel growth control pathway downstream of Fat and Dlish. Using immunoprecipitation and mass spectrometry to search for Dlish partners, we find that Dlish binds the FERM domain growth repressor Expanded (Ex); Dlish SH3 domains directly bind sites in the Ex C terminus. We further show that, in vivo, Dlish reduces the subapical accumulation of Ex, and that loss of Dlish blocks the destabilization of Ex caused by loss of Fat. Moreover, Dlish can bind the F-box E3 ubiquitin ligase Slimb and promote Slimb-mediated ubiquitination of Expanded in vitro. Both the in vitro and in vivo effects of Dlish on Ex require Slimb, strongly suggesting that Dlish destabilizes Ex by helping recruit Slimb-containing E3 ubiquitin ligase complexes to Ex.