Abstract
Citrullinaemia is an inborn error of metabolism resulting from a deficiency of argininosuccinate synthetase. Previous studies of RNA of argininosuccinate synthetase of citrullinaemia patients using S1 nuclease analysis have identified a class of so-called RNA-negative alleles in which no stable mRNA can be detected. To investigate the nature of mutation responsible for such a phenotype, a compound heterozygous citrullinaemia carrying an RNA-negative allele and an allele with a 3′ splice site mutation in intron 6 (IVS6-2A>G) was analysed. Using sequences of a DNA polymorphism and the IVS6-2A>G mutation as markers, approximately equal amounts of pre-mRNAs from allelic genes were detected suggesting that RNA-negative phenotype could not be the result of defect in transcription initiation. A C-to-T transition converting the CGA arginine codon at residue 279 to a TGA termination codon (R279X) was identified by cDNA sequencing. No accumulation of partially spliced pre-mRNAs containing introns immediately upstream and downstream of the nonsense mutation was observed. In addition, no mRNA species of abnormal size was detected when cDNA from the RNA-negative allele was analysed. Hence, there is no indication of nonsense-associated altered splicing (NAS). The most likely event responsible for the RNA-negative phenotype appears to be nonsense-mediated mRNA decay (NMD).
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Acknowledgements
We thank Dr K-B Choo for helpful discussion and critical reading of the manuscript, Drs AL Beaudet and WE O'Brian (Baylor College of Medicine) for providing skin fibroblasts of citrullinaemia patients. This work was supported in part by grants NSC 86-2314-B-075-084 and NSC 87-2314-B-075-089 from the National Science Council, and by a grant from Veterans General Hospital, Taipei.
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Li, CM., Chao, HK., Liu, YF. et al. A nonsense mutation is responsible for the RNA-negative phenotype in human citrullinaemia. Eur J Hum Genet 9, 685–689 (2001). https://doi.org/10.1038/sj.ejhg.5200695
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DOI: https://doi.org/10.1038/sj.ejhg.5200695