Function of a membrane-localized nuclear receptor

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The long-term goals are to understand how a nuclear hormone receptor (NHR)is directed to the plasma membrane in response to a signal and how it might function in a non-nuclear signal transduction system. While the classical view of steroid signaling holds that these hormones bind intracellular receptors that act as ligand-activated transcription factors, extensive evidence indicates that steroids can also act by a non- genomic mechanism at the cell surface. We have identified a C. elegans NHR, DPR-1, that may act through such a membrane-based signaling system, providing the first genetic system for dissecting a membrane- based NHR signaling system. In the presence of a small lipophilic molecule (dauer pheromone or "daumone") DPR-1 moves from the cytoplasm to the plasma membrane. Daumone triggers an alternative, developmental^ arrested state, the dauer larva. A deletion mutation of the dpr-1 gene attenuates responsiveness to dauer pheromone and accelerates exit from the dauer state, and overexpression of DPR- 1 triggers inappropriate dauer development and inhibits exit from the dauer state. The dauer regulatory pathway is required for relocalization of DPR-1 to the membrane. The proposed studies will test the hypothesis that DPR-1 regulates dauer formation through a plasma membrane signal transduction system. Aim 1 will investigate parameters that influence relocalization of DPR-1 in response to dauer pheromone, analyze the essential function of dpr-1, test for redundant partners of DPR-1, and assess the relationship between DPR-1 and the dauer pathway. Aim 2 will identify structural elements and components required for DPR-1 signaling and membrane association, test its signaling function when targeted to the membrane and nucleus, examine the basis for a membrane-tenacious form of DPR-1in dauer larvae, and investigate physical interactions between DPR-1 and other proteins. Aim 3 will examine whether DPR-1responds to dauer pheromone in heterologous cells and whether DPR-1 relatives and the dauer-regulating NHR, DAF- 12, associate with the membrane under dauer-favoring conditions. Aim 4 will initiate RNAi screens to identify components responsible for relocalization of DPR-1and genes with which it collaborates. The elucidation of novel pathways through which NHRs function may advance our understanding of many developmental defects and the hormonal influences on normal human physiology and a variety of human pathologies.