title: Transgenic analysis of rds/peripherin N-glycosylation: effect on dimerization, interaction with rom1, and rescue of the rds null phenotype.
abstract: Rds/peripherin is an integral membrane glycoprotein that is present in the rims of photoreceptor outer segment disks. In mammals, it is thought to stabilize the disk rim through heterophilic interactions with the related nonglycosylated protein roml. Glycosylation of rds/peripherin at asparagine 229 is widely conserved in vertebrates. In this study, we investigated the role of rds/peripherin N-glycosylation. We generated transgenic mice that expressed only S231A-substituted rds/peripherin in their retinas. This protein was not glycosylated but formed covalent dimers with itself and with glycosylated rds/peripherin. Nonglycosylated rds/peripherin also interacted noncovalently with rom1 homodimers to form a heterooligomeric complex. The glycosylated rds/peripherin..rom1 complex bound to concanavalin A-Sepharose, suggesting that the glycan is not directly involved in the interaction between these proteins. In double transgenic mice expressing normal and S231A-substituted rds/peripherin, the mRNA-to-protein ratios were similar for both transgenes, indicating no effect of N-glycosylation on rds/peripherin stability. Finally, expression of nonglycosylated rds/peripherin in transgenic mice rescued the phenotype of outer segment nondevelopment in retinal degeneration slow (rds-/-) null mutants. These observations indicate that N-glycosylation of rds/peripherin is not required for its normal processing, stability, or in vivo function.
authors.affiliation: Center for Basic Neuroscience and Department of Psychiatry, University of Texas Southwestern Medical Center, Dallas 75235-9111, USA.
authors.forename: W
authors.initials: W
authors.lastname: Kedzierski
authors.forename: D
authors.initials: D
authors.lastname: Bok
authors.forename: G H
authors.initials: GH
authors.lastname: Travis
mesh_terms: D000818:Animals; D003208:Concanavalin A; D019281:Dimerization; D005136:Eye Proteins / Q000032:analysis / Q000378:metabolism*; D015870:Gene Expression / Q000502:physiology; D006031:Glycosylation; D015151:Immunoblotting; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008854:Microscopy, Electron; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D064531:Peripherins; D010641:Phenotype; D012162:Retinal Degeneration / Q000378:metabolism; D012374:Rod Cell Outer Segment / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure; D012685:Sepharose; D059470:Tetraspanins; D019076:Transgenes / Q000502:physiology
chemical_list: D005136:Eye Proteins; D007381:Intermediate Filament Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D064531:Peripherins; C578996:Prph2 protein, mouse; C081477:Rom1 protein, mouse; D059470:Tetraspanins; D003208:Concanavalin A; D012685:Sepharose