title: An intramembrane glutamic acid governs peripherin/rds function for photoreceptor disk morphogenesis.
abstract: OBJECTIVE
  Peripherin/rds (P/rds), the product of the retinal degeneration slow (rds) gene, is a tetraspanin protein that plays a pivotal role for photoreceptor outer segment (OS) structure and is involved in a broad spectrum of inherited retinal degenerations. P/rds interacts with the homologous protein rom-1, previously proposed to regulate P/rds function. The authors examined the significance of an intramembrane glutamic acid conserved in all P/rds proteins (and many other tetraspanins) but absent in all rom-1 orthologs.
  
  
  METHODS
  The authors performed isosteric glutamine substitution of the conserved glutamate at position 276, in the fourth transmembrane domain of bovine P/rds, and expressed E276Q P/rds in COS-1 cells and in transgenic mouse photoreceptors of rds +/+, -/+, and -/- backgrounds. Western blot, immunoprecipitation, and sedimentation analyses were used to assess protein structure and interactions. Microscopy and electroretinography were used to characterize transgenic protein localization and retinal photoreceptor structure and function.
  
  
  RESULTS
  E276Q P/rds was expressed, assembled, and properly localized in photoreceptor OSs of transgenic mice. In contrast to wild-type (WT) P/rds, however, this mutant did not rescue the OS structural defects observed in rds -/- and -/+ mice. Moreover, E276Q expression did not prevent the retinal degeneration that occurred as a consequence of OS disruption.
  
  
  CONCLUSIONS
  E276 plays a critical role in P/rds support of photoreceptor OS structure. This finding provides a molecular rationale for asymmetry in P/rds and rom-1 function and for rom-1 regulation of P/rds activity. These findings also suggest that ionizable intramembrane residues may serve regulatory roles for tetraspanin proteins more generally.
authors.affiliation: Eye Research Institute, Oakland University, Rochester, Michigan 48309, USA. goldberg@oakland.edu
authors.forename: Andrew F X
authors.initials: AF
authors.lastname: Goldberg
authors.forename: Linda M
authors.initials: LM
authors.lastname: Ritter
authors.forename: Nidhi
authors.initials: N
authors.lastname: Khattree
authors.forename: Neal S
authors.initials: NS
authors.lastname: Peachey
authors.forename: Robert N
authors.initials: RN
authors.lastname: Fariss
authors.forename: Loan
authors.initials: L
authors.lastname: Dang
authors.forename: Minzhong
authors.initials: M
authors.lastname: Yu
authors.forename: Alyssa R
authors.initials: AR
authors.lastname: Bottrell
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D004596:Electroretinography; D005786:Gene Expression Regulation; D005838:Genotype; D018698:Glutamic Acid / Q000502:physiology*; D047468:Immunoprecipitation; D007381:Intermediate Filament Proteins / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016253:Microscopy, Immunoelectron; D009024:Morphogenesis; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000502:physiology*; D064531:Peripherins; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism* / Q000648:ultrastructure; D012160:Retina / Q000254:growth & development; D012162:Retinal Degeneration / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D019076:Transgenes
chemical_list: D007381:Intermediate Filament Proteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D064531:Peripherins; C578996:Prph2 protein, mouse; D018698:Glutamic Acid