Progressive Rod-Cone Degeneration (PRCD) Protein Requires N-Terminal S-Acylation and Rhodopsin Binding for Photoreceptor Outer Segment Localization and Maintaining Intracellular Stability

Biochemistry. 2016 Sep 13;55(36):5028-37. doi: 10.1021/acs.biochem.6b00489. Epub 2016 Aug 30.

Abstract

The light-sensing outer segments of photoreceptor cells harbor hundreds of flattened membranous discs containing the visual pigment, rhodopsin, and all the proteins necessary for visual signal transduction. PRCD (progressive rod-cone degeneration) protein is one of a few proteins residing specifically in photoreceptor discs, and the only one with completely unknown function. The importance of PRCD is highlighted by its mutations that cause photoreceptor degeneration and blindness in canine and human patients. Here we report that PRCD is S-acylated at its N-terminal cysteine and anchored to the cytosolic surface of disc membranes. We also showed that mutating the S-acylated cysteine to tyrosine, a common cause of blindness in dogs and a mutation found in affected human families, causes PRCD to be completely mislocalized from the photoreceptor outer segment. We next undertook a proteomic search for PRCD-interacting partners in disc membranes and found that it binds rhodopsin. This interaction was confirmed by reciprocal precipitation and co-chromatography experiments. We further demonstrated this interaction to be critically important for supporting the intracellular stability of PRCD, as the knockout of rhodopsin caused a drastic reduction in the photoreceptor content of PRCD. These data reveal the cause of photoreceptor disease in PRCD mutant dogs and implicate rhodopsin to be involved in PRCD's unknown yet essential function in photoreceptors.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acylation
  • Animals
  • Chromatography, Gel
  • Electroporation
  • Eye Proteins / chemistry*
  • Eye Proteins / metabolism
  • Mass Spectrometry
  • Membrane Proteins / chemistry*
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Photoreceptor Cells, Vertebrate / metabolism*
  • Rhodopsin / metabolism*

Substances

  • Eye Proteins
  • Membrane Proteins
  • PRCD protein, mouse
  • Rhodopsin