An outer segment localization signal at the C terminus of the photoreceptor-specific retinol dehydrogenase.
Photoreceptor retinol dehydrogenase (prRDH) is a membrane-associated cytosolic protein that localizes to the outer segments (OS) of rods and cones. Here, we demonstrate that the C-terminal 16 amino acids of prRDH confer membrane association as well as cone and rod OS targeting on a linked green fluorescent protein. Membrane association in transfected 293 cells and in transgenic Xenopus photoreceptors is mediated by fatty acylation at one or more evolutionarily conserved cysteines within the prRDH C-terminal tail. In bovine OS, native prRDH is similarly acylated, and hydrolysis of this linkage releases the protein from the membrane. Efficient OS localization requires both membrane association and the prRDH sequence. (V/I)XPX at the extreme C terminus, which closely resembles the C-terminal sequence that targets opsin/rhodopsin to the OS. Taken together, these data imply that the C-terminal. (V/I)XPX sequence is a general OS localization signal that can function in the context of both integral and peripheral membrane proteins. This strategy for OS localization resembles those used for protein localization to mitochondria, peroxisomes, endosomes, and endoplasmic reticulum; in each case, a short N- or C-terminal sequence is shared among structurally diverse proteins that are targeted to the same subcellular destination.
Luo
Wenqin
W
Department of Neuroscience, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Marsh-Armstrong
Nicholas
N
Rattner
Amir
A
Nathans
Jeremy
J
D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D020816:Amino Acid Motifs / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D002417:Cattle; D002460:Cell Line; D017124:Conserved Sequence / Q000502:physiology; D049452:Green Fluorescent Proteins; D006801:Humans; D007814:Larva / Q000201:enzymology; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D019308:Palmitic Acid / Q000378:metabolism; D010786:Photoreceptor Cells / Q000201:enzymology*; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012374:Rod Cell Outer Segment / Q000201:enzymology*; D014162:Transfection; D014982:Xenopus laevis; D015027:Zebrafish
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D019308:Palmitic Acid; D000429:Alcohol Oxidoreductases; C022520:retinol dehydrogenase