Single base mutation in α5(IV) collagen chain gene converting a conserved cysteine to serine in Alport syndrome

• Published in: Genomics (San Diego, Calif.) Vol. 9; no. 1; pp. 10 - 18
• Main Authors: Zhou, Jing, Barker, David F., Hostikka, Sirkka Liisa, Gregory, Martin C., Atkin, Curtis L., Tryggvason, Karl
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 1991; Elsevier
• Subjects: Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Genes. Genome; Molecular and cellular biology; Molecular genetics; Human; Nephropathy; Restriction fragment length polymorphism; Nucleotide sequence; Collagen; Alport syndrome; Chromosome DNA; Mutation; Secondary structure; Genetic disease
• DOI: 10.1016/0888-7543(91)90215-Z
• ISSN: 0888-7543

We have identified a point mutation in the type IV collagen α5 chain gene ( COL4A5) in Alport syndrome. Variant PstI ( Barker et al., 1990 , Science 248, 1224–1227). and BglII restriction sites with complete linkage with the Alport phenotype have been found in the 3′ end of the COL4A5 gene in the large Utah Kindred P. The approximate location of the variant sites was determined by restriction enzyme mapping, after which this region of the gene (1028 bp) was amplified with the polymerase chain reaction (PCR) from DNA of normal and affected individuals for sequencing analysis. The PCR products showed the absence or presence of the variant PstI and BglII sites in DNA from normal and affected individuals, respectively. DNA sequencing revealed a single base change in exon 3 (from the 3′ end) in DNA from affected individuals, changing the TGT codon of cysteine to the TCT codon for serine. This single base mutation also generated new restriction sites for PstI and BglII. The mutation involves a cysteine residue that has remained conserved in the carboxyl-end noncollagenous domain (NC domain) of all known type IV collagen α chains from Drosophila to man. It is presumably crucial for maintaining the right conformation of the NC domain, which is important for both triple-helix formation and the formation of intermolecular cross-links of type IV collagen molecules.