A presumed missense mutation ofRPGRcauses abnormal RNA splicing with exon skipping

• Published in: American journal of ophthalmology 2004-09, Vol.138 (3), p.504
• Main Authors: Demirci, FYesim K, Radak, Amy L, Rigatti, Brian W, Mah, Tammy S, Gorin, Michael B
• Format: Article
• Language: English
• Subjects: Mutation; Myopia; Proteins
• ISSN: 0002-9394; 1879-1891
• DOI: 10.1016/j.ajo.2004.04.019

Purpose A patient with retinitis pigmentosa demonstrated a novelRPGRmutation (213G>A, last base of exon 2) predicted to cause a missense change (G52R) in the final protein. This study was performed to determine whether this mutation altered the effectiveness of the adjacent splice site. Design Observational case report. Methods Total RNA was extracted from leukocytes of the proband and his carrier mother. Reverse transcription-polymerase chain reaction (RT-PCR) was performed by using the primers flanking exon 2 of RPGR transcript, followed by gel purification and direct sequencing. Results Sequencing revealed skipping of exon 2 in the mutated transcript, leading to in-frame deletion of 42 amino acids affecting the critical RCC1-like domain. Conclusions The last base of exons is conserved as "G" in 80% of splicing consensus sequences, yet when changed, can completely disrupt constitutive splicing as in this patient. Our data confirm that the evaluation of the effects of some DNA sequence alterations at the RNA level might have important implications for appropriate genotype-phenotype correlations.