Coupling of Human Rhodopsin to a Yeast Signaling Pathway Enables Characterization of Mutations Associated with Retinal Disease

摘要: G protein-coupled receptors (GPCRs) are crucial sensors of extracellular signals in eukaryotes, with multiple GPCR mutations linked to human diseases. With the growing number sequenced genomes, determining pathogenicity a mutation is challenging, but can be aided by direct measurement GPCR-mediated signaling. This particularly difficult for visual pigment rhodopsin-a activated light-for which hundreds have been inherited degenerative retinal diseases such as retinitis pigmentosa. In this study, we successfully engineered, first time, activation rhodopsin yeast mating pathway, resulting signaling via fluorescent reporter. We combine novel assay light-dependent studies subcellular localization, and upregulation unfolded protein response misfolded protein. use these assays characterize panel known molecular phenotypes, finding that maintains similar phenotype yeast, some interesting differences. Furthermore, compare our clinical phenotypes from patients disease-linked mutations. demonstrate engineered strain useful mutant classification, helping determine mechanisms underlying their pathogenicity. approach may also applied better understand relevance other mutations, furthering tool investigating relevant disease.