A T3 allele in the CFTR gene exacerbates exon 9 skipping in vas deferens and epididymal cell lines and is associated with congenital bilateral absence of vas deferens (CBAVD)
A T3 allele in the CFTR gene exacerbates exon 9 skipping in vas deferens and epididymal cell lines and is associated with congenital bilateral absence of vas deferens (CBAVD)
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DOI：
10.1002/humu.20115
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发表时间：
2005-01-01
期刊：
影响因子：
3.9
通讯作者：
Tuffery-Giraud, S
中科院分区：
文献类型：
--
作者：
Disset, A;Michot, C;Tuffery-Giraud, S
人CFTR内含子8 3'端（TG）m（T）n多态性基因座上的不同等位基因决定了外显子9的剪接效率。我们在一例先天性双侧输精管缺如（CBAVD）患者中发现了一种新的TG 12 T3等位基因，该患者在另一条染色体上携带[TG 11 T7; p.Phe508Cys; p.Met470Va1]单倍型。为了更好地理解外显子9剪接的复杂调控，我们分析了来自肺、结肠、睾丸、输精管和附睾的6个CFTR表达上皮细胞系中正确剪接的CFTR转录物的水平，这些细胞系瞬时转染了4个CFTR小基因（pTG 11 T7、pTG 12 T7、pTG 12 T5和pTG 12 T3）。在这项工作中，我们表明，减少Ts在多态性位点的TG 12背景决定了一个细胞，类型依赖性减少外显子9+转录本是不相关的基础剪接效率的细胞系。这些数据强调了T5等位基因在CBAVD中的作用，并将T3等位基因鉴定为严重囊性纤维化（CF）致病突变。最后，紫外线交联实验表明，组织特异性反式作用剪接因子不有助于外显子9剪接的细胞系之间发现的不同模式。然而，我们观察到较低数量的Ts可以以组织特异性方式改变TDP-43（TDP 43或TARDBP）与其特异性靶ug 12的结合。我们的研究结果支持这样的想法，即一般剪接因子的比例在外显子9选择性剪接的组织变异性中起着重要作用。(C)2004 Wiley-Liss，Inc.
The different alleles at the (TG)m(T)n polymorphic loci at the 3' end of the human CFTR intron 8 determine the efficiency by which exon 9 is spliced. We identified a novel TG12T3 allele in a congenital bilateral absence of vas deferens (CBAVD) patient who carries a [TG11T7; p.Phe508Cys; p.Met470Va1] haplotype on the other chromosome. To better understand the complex regulation of exon 9 splicing, we analyzed the levels of correctly spliced CFTR transcripts in six CFTR-expressing epithelial cell lines derived from lung, colon, testis, vas deferens, and epididymis transiently transfected with four CFTR minigenes (pTG11T7, pTG12T7, pTG12T5, and pTG12T3). In this work, we show that a decrease in the Ts at the polymorphic locus in a TG12 background determines a cell,type dependent reduction in exon 9+ transcripts that is not related to the basal splicing efficiency in the cell line. These data emphasize the role of the T5 allele in CBAVD and identify the T3 allele as a severe cystic fibrosis (CF) disease-causing mutation. Finally, UV cross-linking experiments demonstrated that tissue-specific trans-acting splicing factors do not contribute to the different patterns of exon 9 splicing found between the cell lines. However, we observed that lower numbers of Ts can alter the binding of TDP-43 (TDP43 or TARDBP) to its specific target ug12 in a tissue-specific manner. Our results support the idea that the ratio of general splicing factors plays a role in the tissue variability of exon 9 alternative splicing. (C) 2004 Wiley-Liss, Inc.