Abstract
Abstract: :
Purpose: To identify genes that interact with NR2E3 to regulate cone photoreceptor development and function. Methods: The rd7/rd7 mouse, harboring a mutation in Nr2e3, was used to study cone photoreceptor development and function. Electrophoretic mobility shift assay (EMSA) was performed using NR2E3 protein expressed in bacteria and 32P labeled oligonucleotides of putative response element sequence upstream of genes misregulated in rd7/rd7mice. Yeast 2–hybrid analysis was performed using full–length NR2E3 and a construct containing only the ligand–binding and dimerization domain of NR2E3 as bait. Interacting factors were derived from brain and retina prey libraries. Results: Previously, real time PCR and subtractive hybridization analyses identified 16 genes misregulated in rd7/rd7 mice. This study shows several of these genes including rpe65, gpr91, gnat2, nrbf2, and trß2 contain 5' upstream target sequences that NR2E3 binds. Yeast 2–hybrid analysis identified factors that directly interact with NR2E3. Conclusions:Genes that NR2E3 regulates and those that NR2E3 recruits as co–suppressors or co–activators may provide entry points for pathways in which NR2E3 functions during cone photoreceptor development and maintenance, and potentially novel pathways for visual transduction in the adult retina.
Keywords: photoreceptors • transcription factors • retinal development