Department of Medical Biochemistry, Oslo University Hospital, PO Box 4950 Nydalen, N-0424 Oslo, Norway.
Mol Genet Metab. 2011 Nov;104(3):289-94. doi: 10.1016/j.ymgme.2011.07.012. Epub 2011 Jul 20.
Mucopolysaccharidosis type I is an autosomal recessive disorder caused by deficiency of α-l-iduronidase, encoded by the IDUA gene. More than 100 disease causing mutations have been reported in the gene, resulting in a wide range of phenotypes. Here we describe a previously unreported IDUA splice site mutation (NG_008103.1:g.21632G>C; NM_000203.3:c.1727+3G>C) causing a Hurler phenotype in a patient heterozygous for the common p.Q70X (NG_008103.1:g.5862C>T) mutation. Sequence analysis of IDUA transcripts demonstrated that the g.21632G>C mutation results in aberrant splicing of intron 12 (NM_000203.3:c.1727_1728insGTCC), introducing a frame shift and premature termination codon (NP_000194.2:p.Cys577SerfsX15). Gene expression studies suggest that the deleterious effect of the mutation is primarily due to a C-terminal truncation of the encoded polypeptide. Furthermore, we observed that both normal and mutant IDUA alleles give rise to alternatively spliced transcripts in leukocytes. Exclusion of exon 4 appeared to be the predominant alternative splicing event, probably resulting in polypeptides lacking iduronidase activity. The Hurler patient demonstrated exon 4 skipping in 5.6% of IDUA transcripts, while exon 4 skipping ranged 25-34% of transcripts among healthy individuals (n=5). Alternative splicing might represent a mechanism for regulation of this enzyme, and the lower level of exon 4 skipping in the patient might be a response to intracellular accumulation of iduronidase substrates. Molecular characterization of IDUA mutations and splicing may assist early prediction of mucopolysaccharidosis type I phenotypes and increase the understanding of disease mechanisms. This is important considering the choice of current treatment options and for the development of future therapies.
黏多糖贮积症 I 型是一种常染色体隐性遗传病,由 IDUA 基因编码的α-L-艾杜糖苷酸酶缺乏引起。该基因已报道超过 100 种致病突变,导致表型广泛。在此,我们描述了一个以前未报道的 IDUA 剪接位点突变(NG_008103.1:g.21632G>C;NM_000203.3:c.1727+3G>C),导致一位携带常见 p.Q70X(NG_008103.1:g.5862C>T)突变的杂合子患者表现为 Hurler 表型。IDUA 转录本的序列分析表明,g.21632G>C 突变导致内含子 12 的异常剪接(NM_000203.3:c.1727_1728insGTCC),引入移码并提前终止密码子(NP_000194.2:p.Cys577SerfsX15)。基因表达研究表明,该突变的有害影响主要是由于编码多肽的 C 端截短。此外,我们观察到白细胞中正常和突变的 IDUA 等位基因都产生了选择性剪接的转录本。外显子 4 的缺失似乎是主要的选择性剪接事件,可能导致缺乏艾杜糖苷酸酶活性的多肽。Hurler 患者的 IDUA 转录本中存在 5.6%的外显子 4 缺失,而健康个体的转录本中外显子 4 缺失范围为 25-34%(n=5)。选择性剪接可能是该酶调控的一种机制,患者中外显子 4 缺失的水平较低可能是对细胞内艾杜糖苷酸酶底物积累的一种反应。IDUA 突变和剪接的分子特征可能有助于早期预测黏多糖贮积症 I 型表型,并增加对疾病机制的理解。考虑到当前治疗选择的选择和未来治疗的发展,这一点非常重要。