ADVIRC is caused by distinct mutations in BEST1 that alter pre-mRNA splicing
ADVIRC is caused by distinct mutations in BEST1 that alter pre-mRNA splicing
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DOI：
10.1136/jmg.2008.059881
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发表时间：
2009-09-01
影响因子：
4
通讯作者：
Manson, F. D. C.
中科院分区：
文献类型：
--
作者：
Burgess, R.;MacLaren, R. E.;Manson, F. D. C.
常染色体显性玻璃体视网膜脉络膜病（ADVIRC）是一种视网膜营养不良症，常与青光眼和白内障相关，是“视网膜病变”表型谱的一部分。先前已经证明，ADVIRC是由BEST1突变引起外显子跳变并导致产生缩短和内部缺失的同工型引起的。本研究描述了一种新的ADVIRC突变，并表明它破坏了一个外显子剪接增强子（ESE）位点，改变了剪接相关SR蛋白的结合。与之前的ADVIRC突变一样，在离体剪接实验中，外显子6中的新型c. 704T -> c突变改变了正常的剪接。与附近与Best病相关的外显子6突变（C . 703g -> C）相比，该突变和另一个引起advirc的外显子6突变（C . 707g -> A）在es依赖剪接试验中减弱或消除了剪接。采用包含ADVIRC和Best疾病突变的RNA寡核苷酸和四种最常研究的SR蛋白进行凝胶移位测定。尽管SC35、SRp40和SRp55蛋白与野生型和突变序列的结合强度相似，但与野生型或最佳疾病突变序列相比，ASF/SF2与两种advirc突变序列的结合强度增加。这两个外显子6 ADVIRC突变的外显子跳变及其对ASF/SF2的亲和力表明，包含这些突变的区域可能是剪接复合外显子调控元件（CERES）位点的一部分。
Autosomal dominant vitreoretinochoroidopathy (ADVIRC), a retinal dystrophy often associated with glaucoma and cataract, forms part of a phenotypic spectrum of 'bestrophinopathies'. It has been shown previously that ADVIRC results from BEST1 mutations that cause exon skipping and lead to the production of shortened and internally deleted isoforms. This study describes a novel ADVIRC mutation and show that it disrupts an exonic splice enhancer (ESE) site, altering the binding of a splicing-associated SR protein. As with previous ADVIRC mutations, the novel c. 704T -> C mutation in exon 6 altered normal splicing in an ex vivo splicing assay. Both this and another exon 6 ADVIRC-causing mutation (c.707G -> A) either weakened or abolished splicing in an ESE-dependent splice assay compared with a nearby exon 6 mutation associated with Best disease (c.703G -> C). Gel shift assays were undertaken with RNA oligonucleotides encompassing the ADVIRC and Best disease mutations with four of the most commonly investigated SR proteins. Although SC35, SRp40 and SRp55 proteins all bound to the wild-type and mutated sequences with similar intensities, there was increased binding of ASF/SF2 to the two ADVIRC-mutated sequences compared with the wild-type or Best disease-mutated sequences. The exon skipping seen for these two exon 6 ADVIRC mutations and their affinity for ASF/SF2 suggests that the region encompassing these mutations may form part of a CERES (composite exonic regulatory elements of splicing) site.