ABERRANT SPLICING OF THE COL4A5 GENE IN PATIENTS WITH ALPORT SYNDROME

A variety of mutations have been identified in the X-linked type IV co llagen alpha 5 chain (COL4A5) gene in patients with Alport syndrome. A substantial number of these mutations were predicted to have an effec t on RNA splicing. For 4 such mutations in our group of patients the e ffect of the DNA mutation on the COL4A5 mRNA structure and stability w as analysed. An alteration of the invariant splice acceptor site of in tron 41 resulted in a shift of the actual splicing to either a cryptic splice site within exon 42 or the normal splice site in the next intr on. A single base substitution of the final nucleotide of exon 48 resu lted in the removal of the entire exon. Two frameshift mutations, a 10 basepair duplication in exon 49 and a single base deletion in exon 41 , were incorporated in the mRNA as such and resulted in a stretch of m issense codons terminated by a premature stop codon. Exon skipping was occasionally observed in these samples, but not reproducibly in every experiment. In healthy controls exon skipping was never detected. Ana lysis of female carriers revealed that in only one case was the stabil ity of the mutated mRNA reduced in comparison with the normal transcri pt. The extent to which the noncollagenous domain was predicted to be deleted correlated with the severeness of the disease.