Molecular studies of hypoxanthine-guanine phosphoribosyltransferase mutations in six Australian families.

Genomic deoxyribonucleic acid (DNA) was isolated from six hemizygotes and five heterozygotes from unrelated families exhibiting the full clinical spectrum of hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency. The DNA was digested with the restriction endonucleases, Bam H1, Pst 1 and Taq 1, previously found to be useful in demonstrating restriction fragment length polymorphism (RFLP) at the HPRT locus of the X-chromosome. DNA blotting experiments using a full length HPRT-cDNA probe, have revealed RFLPs in three families which may prove useful for the diagnosis of HPRT deficiency and the determination of heterozygosity. Total ribonucleic acid (RNA) was also extracted from our 11 subjects and analysed by Northern blotting for the presence of HPRT-messenger (mRNA). Apparently normal HPRT-mRNA was demonstrated in all the hemizygotes and heterozygotes for partial HPRT deficiency. In the families with complete HPRT deficiency (Lesch-Nyhan syndrome), the heterozygotes had normal HPRT-mRNA. However, one hemizygote had a complete absence of message for HPRT, while the other hemizygote had considerably reduced amounts of this message