The inturned (in) gene is a component of the frizzled (fz) signaling pathway that controls the polarity of hairs and bristles in the epidermis of Drosophila. It appears to act downstream of fz, which encodes a putative receptor for a tissue polarity signal. The in gene encodes a novel protein that had been suggested to contain two potential transmembrane domains. It has been suggested that the In protein interacts with the actin cytoskeleton to regulate the formation of the pupal wing prehairs that become adult hairs. The initiation of prehairs is normally restricted to the vicinity of the distal most vertex along the apical surface of the pupal wing cells. In an in mutant, prehairs initate at a variety of locations along the apical cell periphery. We have used immunofluorescence to study the subcellular localization of the In protein. When expressed in cultured cells, we found that In is a cytoplasmic protein. However, we found that it is localized in the vicinity of plasma membrane and the cortical actin cytoskeleton of Drosophila wing disc and pupal wing cells. Thus, in wing cells the In protein is localized to the region of the cell where it appears to function. This subcellular localization presumably requires the function of other proteins and may represent a regulatory mechanism. Our data suggest that fz does not play a major role in the subcellular localization of In. The In protein is notably insoluble in buffers containing high salt and nonionic detergents. This lack of solubility is significantly reduced in fz and mwh mutants, implying that it may be related to the mechanism of in function.
Copyright 1999 Wiley-Liss, Inc.