COL4A5 splice site mutation and α5(IV) collagen mRNA in Alport syndrome
| Autor: | Jing Zhou, Oliver Pullig, K O Netzer, Karl Tryggvason, Ulrich Frei, Manfred Weber |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Adult
Male Sequence analysis RNA Splicing DNA Mutational Analysis Molecular Sequence Data Nephritis Hereditary Biology urologic and male genital diseases Polymerase Chain Reaction Exon Type IV collagen medicine otorhinolaryngologic diseases Humans RNA Messenger Alport syndrome Genetics Splice site mutation Base Sequence Point mutation Intron DNA medicine.disease Molecular biology Transplantation Nephrology Collagen |
| Zdroj: | Kidney International. 43(2):486-492 |
| ISSN: | 0085-2538 |
| DOI: | 10.1038/ki.1993.71 |
| Popis: | COL4A5 splice site mutation and α5(IV) collagen mRNA in Alport syndrome. Mutations affecting the COL4A5 gene encoding the α5 chain of type IV collagen, are involved in the pathogenesis of X-linked Alport syndrome. We used denaturing gradient gel electrophoresis (DGGE) to screen PCR amplified exons of COL4A5 for point mutations in a set of 18 Alport patients previously characterized by Southern blotting. One sequence variant was identified in the exon 38 region of a male Alport patient. Sequence analysis revealed a G to C trans version in the 5′ intron splice donor site downstream from exon 38 (GT to CT). To determine the effect of the mutation on mRNA splicing, α5(IV) cDNA was generated from total RNA of peripheral blood lymphocytes. Subsequent cDNA PCR yielded a product 81 base pairs shorter in the affected Alport patient, compared to normal controls. The absence of exon 38 from the α5(IV) cDNA was confirmed by sequence analysis. The results demonstrated that the mutation leads to skipping of exon 38 in the processing of α5(IV) pre-mRNA. The shortened transcript lacked 27 codons encoding a Gly-X-Y-repeat sequence with a preserved reading frame, enabling the translation of codons further downstream. Clinically, the patient presented with juvenile onset Alport syndrome, end-stage renal failure, and deafness. He had no ocular lesions. Typical ultrastructural changes of the glomerular basement membrane (GBM) were shown on electron microscopy. The patient developed anti-GBM antibodies after renal transplantation, however, renal function deteriorated only moderately. |
| Databáze: | OpenAIRE |
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