Aggregation of the constitutively active K296E rhodopsin mutant contributes to retinal degeneration.

Abstract

A K296E mutation in rhodopsin causes autosomal dominant retinitis pigmentosa, a progressive retinal degenerative disease. Early in vitro characterizations of this mutation studied on a bovine rhodopsin background indicated that the mutation causes the receptor to be constitutively active. This molecular defect has been the primary focus when considering the pathogenic mechanism of the mutation. Knockin mice expressing the K296E rhodopsin mutant were generated and characterized to better understand the pathogenic mechanism of the mutation. Knockin mice exhibited progressive retinal degeneration characteristic of retinitis pigmentosa. The K296E rhodopsin mutant mislocalized in photoreceptor cells and, surprisingly, appeared to aggregate, as indicated by the dye PROTEOSTAT, which binds protein aggregates. The propensity of the K296E rhodopsin mutant to aggregate was tested and confirmed in vitro but was dependent on the species background of rhodopsin. The K296E mutation on either murine or human rhodopsin backgrounds exhibited similar propensities to aggregate. The same mutation on a bovine rhodopsin background, however, exhibited a lower propensity to aggregate, indicating this species background does not adequately model the effects of the K296E mutation. In contrast to previous expectations, we demonstrate here that aggregation of the K296E rhodopsin mutant can promote photoreceptor cell loss.