The C terminus of peripherin/rds participates in rod outer segment targeting and alignment of disk incisures.
Protein targeting is essential for domain specialization in polarized cells. In photoreceptors, three distinct membrane domains exist in the outer segment: plasma membrane, disk lamella, and disk rim. Peripherin/retinal degeneration slow (rds) and rom-1 are photoreceptor-specific members of the transmembrane 4 superfamily of transmembrane proteins, which participate in disk morphogenesis and localize to rod outer segment (ROS) disk rims. We examined the role of their C termini in targeting by generating transgenic Xenopus laevis expressing green fluorescent protein (GFP) fusion proteins. A GFP fusion containing residues 317-336 of peripherin/rds localized uniformly to disk membranes. A longer fusion (residues 307-346) also localized to the ROS but exhibited higher affinity for disk rims than disk lamella. In contrast, the rom-1 C terminus did not promote ROS localization. The GFP-peripherin/rds fusion proteins did not immunoprecipitate with peripherin/rds or rom-1, suggesting this region does not form intermolecular interactions and is not involved in subunit assembly. Presence of GFP-peripherin/rds fusions correlated with disrupted incisures, disordered ROS tips, and membrane whorls. These abnormalities may reflect competition of the fusion proteins for other proteins that interact with peripherin/rds. This work describes novel roles for the C terminus of peripherin/rds in targeting and maintaining ROS structure and its potential involvement in inherited retinal degenerations.
Tam
Beatrice M
BM
Department of Neuroscience, University of Connecticut Health Center, Farmington, Connecticut 06030, USA. btam@interchange.ubc.ca
Moritz
Orson L
OL
Papermaster
David S
DS
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002417:Cattle; D002455:Cell Division; D049452:Green Fluorescent Proteins; D006801:Humans; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000737:chemistry*; D008164:Luminescent Proteins / Q000737:chemistry / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000737:chemistry*; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D064531:Peripherins; D017434:Protein Structure, Tertiary; D017382:Reactive Oxygen Species; D012374:Rod Cell Outer Segment / Q000378:metabolism*; D013997:Time Factors; D019076:Transgenes; D029867:Xenopus Proteins; D014982:Xenopus laevis
D007381:Intermediate Filament Proteins; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; C578995:PRPH protein, Xenopus; C578994:PRPH protein, human; C578997:PRPH2 protein, human; D064531:Peripherins; D017382:Reactive Oxygen Species; D029867:Xenopus Proteins; D049452:Green Fluorescent Proteins