A deep intronic mutation in the ankyrin-1 gene causes diminished protein expression resulting in hemolytic anemia in mice.
A deep intronic mutation in the ankyrin-1 gene causes diminished protein expression resulting in hemolytic anemia in mice.
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DOI：
10.1534/g3.113.007013
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发表时间：
2013-10-03
期刊：
影响因子：
--
通讯作者：
Du X
中科院分区：
文献类型：
--
作者：
Huang H;Zhao P;Arimatsu K;Tabeta K;Yamazaki K;Krieg L;Fu E;Zhang T;Du X
跨膜蛋白和基于血影蛋白的细胞骨架之间的连接对于红细胞的膜弹性是必需的。介导这种连接的蛋白质的突变导致各种类型的溶血性贫血。在这里，我们报告了一种新的N-乙基-N-亚硝基脲诱导的锚蛋白-1突变，命名为hema 6，它通过蛋白表达的轻度减少引起小鼠遗传性球形红细胞增多症。致病突变被追溯到位于Ank 1基因内含子13深处的单核苷酸转换。在体外小基因剪接试验发现两个异常剪接的转录本含有隐蔽外显子的片段Ank 1内含子13。包含隐蔽外显子引入了提前终止密码子，其导致突变体转录物在体内的无义介导的衰变。因此，在纯合子小鼠中，仅表达野生型锚蛋白-1，尽管表达水平为野生型小鼠的70%。杂合子表现出类似的遗传性球形红细胞表型，源于中间蛋白质表达水平，表明突变的单倍不足。在突变小鼠中观察到完整的跨膜蛋白，带3，和潜在的细胞骨架之间的联系减弱，这是由于锚蛋白-1提供的高亲和力结合位点减少。Hema 6是Ank 1等位基因系列中唯一已知的以降低水平表达全长典型锚蛋白-1的小鼠突变体，这一事实使得它对于研究锚蛋白-1数量缺陷的功能影响特别有用。
Linkage between transmembrane proteins and the spectrin-based cytoskeleton is necessary for membrane elasticity of red blood cells. Mutations of the proteins that mediate this linkage result in various types of hemolytic anemia. Here we report a novel N-ethyl-N-nitrosourea−induced mutation of ankyrin-1, named hema6, which causes hereditary spherocytosis in mice through a mild reduction of protein expression. The causal mutation was traced to a single nucleotide transition located deep into intron 13 of gene Ank1. In vitro minigene splicing assay revealed two abnormally spliced transcripts containing cryptic exons from fragments of Ank1 intron 13. The inclusion of cryptic exons introduced a premature termination codon, which leads to nonsense-mediated decay of the mutant transcripts in vivo. Hence, in homozygous mice, only wild-type ankyrin-1 is expressed, albeit at 70% of the level in wild-type mice. Heterozygotes display a similar hereditary spherocytosis phenotype stemming from intermediate protein expression level, indicating the haploinsufficiency of the mutation. Weakened linkage between integral transmembrane protein, band 3, and underlying cytoskeleton was observed in mutant mice as the result of reduced high-affinity binding sites provided by ankyrin-1. Hema6 is the only known mouse mutant of Ank1 allelic series that expresses full-length canonical ankyrin-1 at a reduced level, a fact that makes it particularly useful to study the functional impact of ankyrin-1 quantitative deficiency.
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