独立性无症状甘油激酶缺乏症伴突变体RNA的接受点突变和无意义介导衰变

摘要

Isolated glycerol kinase deficiency (GKD) is an X-linked inborn error of metabolism that is either symptomatic or asymptomatic.GKD is due to deletions of,or mutations within,the GK gene,and there is no genotype-phenotype correlation. We identified three patients with asymptomatic GKD,determined that they had GK splice-site mutations,and studied the stability of their GK mRNA to understand the molecular mechanism of the GKD. All three patient mutations caused a frameshift and introduction of a premature stop codon. A fourth patient hadan Alu insertion in intron 4 that led to alternative splicing. To study the effect of splice-site mutations on RNA species,we performed reverse transcriptase PCR and found only normalsized products for all patients. Incubation with anisomycin to block nonsense-mediated decay (NMD),revealed two RNA species for each individual. Sequence analysis revealed that the larger bands represented the wild-type GK RNA and smaller bands represented mutant misspliced RNA,suggesting that the abnormal RNA species were targeted by NMD. Normal RNA species observed in each patient are likely responsible for their mild phenotypes. We speculate that influences on RNA processing and protein stability represent modifiers of the GKD phenoty-