G to T transversion at position +5 of a splice donor site causes skipping of the preceding exon in the type III procollagen transcripts of a patient with Ehlers-Danlos syndrome type IV.
G to T transversion at position +5 of a splice donor site causes skipping of the preceding exon in the type III procollagen transcripts of a patient with Ehlers-Danlos syndrome type IV.
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DOI：
10.1016/s0021-9258(19)67780-x
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发表时间：
1991-03
期刊：
影响因子：
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通讯作者：
Brendan Lee;Emilia VitaleS;Andrea Superti-Furgall;Beat Steinmannll;Francesco Ramirez
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作者：
Brendan Lee;Emilia VitaleS;Andrea Superti-Furgall;Beat Steinmannll;Francesco Ramirez
我们在ehers - danlos综合征IV型患者中发现了剪接突变，这是一种与III型胶原蛋白功能障碍相关的遗传性结缔组织疾病。该突变首先定位于患者的III型前胶原mRNA，方法是利用聚合酶链反应扩增几个重叠片段的逆转录产物。扩增产物在24-26外显子序列上显示两个不同的片段，一个是正常大小，另一个是缺乏25外显子的99个碱基对。扩增的基因组产物测序发现，在患者的一个前胶原III基因中，内含子25剪接供体位点+5位置发生了G到T的翻转。等位基因构建的表达将T代替G与25外显子序列的跳跃联系起来。就像之前其他胶原蛋白基因的剪接突变一样，降低培养成纤维细胞的温度几乎消除了外显子跳变。作为本研究的一部分，我们还鉴定了一个高度多态性的内含子DNA序列，其不同的等位基因形式可以很容易地通过聚合酶链反应技术检测到。
We identified a splicing mutation in a patient with Ehlers-Danlos syndrome type IV, a heritable connective tissue disorder associated with dysfunctions of type III collagen. The mutation was first localized in the patient's type III procollagen mRNA by amplifying the reverse transcribed product in several overlapping fragments using the polymerase chain reaction. Amplified products spanning exon 24-26 sequences displayed two distinct fragments, one of normal size and the other lacking the 99 base pairs of exon 25. Sequencing of amplified genomic products identified a G to T transversion at position +5 of the splice donor site of intron 25 in one of the patient's procollagen III genes. Expression of allelic minigene constructs correlated the T for G substitution with skipping of exon 25 sequences. Like previously characterized splicing mutations in other collagen genes, lowering the temperature at which the patient's fibroblasts were incubated nearly abolished exon skipping. As a part of this study, we also identified a highly polymorphic, intronic DNA sequence whose different allelic forms can be detected easily by the polymerase chain reaction technique.