Identification of a 2 Mb Human Ortholog of Drosophila eyes shut/spacemaker that Is Mutated in Patients with Retinitis Pigmentosa

In patients with autosomal-recessive retinitis pigmentosa (arRP), homozygosity mapping was performed for detection of regions harboring genes that might be causative for RP In one affected sib pair, a shared homozygous region of 5.0 Mb was identified oil chromosome 6, within the RP25 locus. One of the genes residing in this interval was the retina-expressed gene EGFL 11. Several genes resembling EGFL I I were predicted just centromeric of EGFL 11. Extensive long-range RT-PCR, combined with 5'- and 3'- RACE analysis, resulted it the identification of a 10-kb transcript, starting with the annotated exons of EGFL I I and spanning 44 exons and 2 Mb of genomic DNA. The transcript is predicted to encode a 31 65-aa extracellular protein containing 28 EGF-like and five laminin A G-like domains. Interestingly, the second part of the protein was found to be the human ortholog of Drosophila eyes shut (cys), also known as spacemaker, a protein essential for photoreceptor morphology. Mutation analysis in the sib pair homozygous at RP25 revealed a nonsense mutation (p.Tyr3156X) segregating with RP The same mutation was identified homozygously in three arRP siblings of an unrelated family. A frame-shift Mutation (pPro2238ProfsX16) was found in an isolated RP patient. fir conclusion, we identified a gene, coined eyes shut homolog (EYS), consisting of EGFL11 and the human ortholog of Drosophila eys, which is mutated in patients with arRP. With a size of 2 Mb, it is one of the largest human genes, and it is by far the largest retinal dystrophy gene. The discovery of EYS might shed light on a critical component of photoreceptor morphogenesis.