The tetraspanin disc proteins, peripherin-2 and ROM1, facilitate CNG channel localization to the rod outer segment

ABSTRACT The light-responsive outer segment of rod photoreceptors is composed of two distinct membrane subdomains: discs and the plasma membrane. We investigate how the disc protein peripherin-2 is engaged in CNG channel delivery to the outer segment. Instead of forming outer segments, peripherin-2 knockout ( Rds -/- ) photoreceptors release ciliary ectosomes, in which CNG channel levels are markedly reduced relative to other outer segment proteins. This is intriguing as downregulation of the CNG channel is not a general feature of degenerative mouse models with dysmorphic outer segments. Overexpression of the β1-subunit of CNG in Rds -/- rods reveals that the majority is trapped in intracellular membranes, but is restored to the outer segment by co-expressing peripherin-2. We test peripherin-2 chimeras containing either the N-terminus, tetraspanin core, or C-terminus and find that the tetraspanin domain is sufficient to localize CNGβ1 to the outer segment. We further show that the membrane remodeling function of the tetraspanin domain facilitates this process by the redundant action of the tetraspanin domain from ROM1 and the reemergence of the endogenous CNG channel in aged Rds -/- rods that have produced ciliary membrane protrusions.