The C terminus of peripherin/rds participates in rod outer segment targeting and alignment of disk incisures

Mol Biol Cell. 2004 Apr;15(4):2027-37. doi: 10.1091/mbc.e03-09-0650. Epub 2004 Feb 6.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Cattle
  • Cell Division
  • Green Fluorescent Proteins
  • Humans
  • Immunohistochemistry
  • Intermediate Filament Proteins / chemistry*
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / metabolism
  • Membrane Glycoproteins / chemistry*
  • Microscopy, Confocal
  • Microscopy, Electron
  • Microscopy, Immunoelectron
  • Molecular Sequence Data
  • Nerve Tissue Proteins / chemistry*
  • Peripherins
  • Protein Structure, Tertiary
  • Reactive Oxygen Species
  • Rod Cell Outer Segment / metabolism*
  • Time Factors
  • Transgenes
  • Xenopus Proteins
  • Xenopus laevis

Substances

  • Intermediate Filament Proteins
  • Luminescent Proteins
  • Membrane Glycoproteins
  • Nerve Tissue Proteins
  • PRPH protein, Xenopus
  • PRPH protein, human
  • PRPH2 protein, human
  • Peripherins
  • Reactive Oxygen Species
  • Xenopus Proteins
  • Green Fluorescent Proteins