Enhanced exon 2 skipping caused by c.910G>A variant and alternative splicing of MEFV genes in two independent cases of familial Mediterranean fever
Enhanced exon 2 skipping caused by c.910G>A variant and alternative splicing of MEFV genes in two independent cases of familial Mediterranean fever
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在两例独立的家族性地中海热病例中，c.910G>A 变异和 MEFV 基因的选择性剪接导致外显子 2 跳跃增强
DOI：
10.1007/s10165-011-0461-4
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发表时间：
2012
期刊：
影响因子：
2.2
通讯作者：
Yachie A
中科院分区：
文献类型：
--
作者：
Tone Y;Toma T;Toga A;Sakakibara Y;Wada T;Yabe M;Kusafuka H;Yachie A
大多数报告的家族性地中海热（FMF）病例涉及mefv的错义突变集中在外显子10。我们经历了mefvgene中一个独特变体的两个独立谱系，该变体可能由于增强的选择性剪接而导致过多的外显子2跳变。在这项研究中，我们试图阐明mefv变异体作为FMF表型原因的分子机制。从志愿者和两名携带mefv基因纯合子c.910G . >A变体的患者中提取外周血。分别利用外显子1和外显子3的正向引物和反向引物比较mefv信使RNA （mRNA）在单核细胞和粒细胞中的表达模式。通过转染野生型和变异型mefv基因到HEK293T细胞中检测pyrin的表达谱。正常大小的mRNA的表达在这些患者中极度减少，而异常短的mRNA的表达，删除外显子2 （Δex2），显著增加。免疫组织化学和免疫印迹分析显示，在转染Δex2 cDNA的细胞中有一个截断的免疫反应性pyrin蛋白。mefvgene c.910G>A变异导致异常剪接加速，蛋白质大小异常，可能导致异常pyrin功能。这是第一个报告表明，除错义突变外，mefvvariant是导致FMF表型的原因。
Most reported cases of familial Mediterranean fever (FMF) involve missense mutations ofMEFVconcentrated within exon 10. We experienced two independent pedigrees of a unique variant in theMEFVgene that might cause excessive exon 2 skipping due to enhanced alternative splicing. In this study, we tried to elucidate the molecular mechanism of theMEFVvariant as a cause of the FMF phenotype. Peripheral blood was obtained from volunteers and two patients with homozygous c.910G>A variant of theMEFVgene.MEFVmessenger RNA (mRNA) expression patterns in mononuclear cells and granulocytes were compared using forward and reverse primers from exons 1 and 3, respectively. Expression profiles of pyrin were examined by transfecting wild-type and variantMEFVgenes into HEK293T cells. Expression of normal-sized mRNA was extremely reduced in these patients, whereas that of aberrant short mRNA, deleting exon 2 (Δex2), was significantly increased. Immunohistochemical and immunoblotting analyses revealed a truncated immunoreactive pyrin protein in cells transfected with Δex2 cDNA. TheMEFVgene c.910G>A variant results in accelerated aberrant splicing with abnormal protein size, presumably leading to anomalous pyrin function. This is the first report to show that anMEFVvariant other than missense mutation is responsible for the FMF phenotype.