Disease mechanism for retinitis pigmentosa (RP11) caused by mutations in the splicing factor gene PRPF31
Disease mechanism for retinitis pigmentosa (RP11) caused by mutations in the splicing factor gene PRPF31
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DOI：
10.1093/hmg/11.25.3209
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发表时间：
2002-12-01
影响因子：
3.5
通讯作者：
Wilkie, SE
中科院分区：
文献类型：
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作者：
Deery, EC;Vithana, EN;Wilkie, SE
本研究调查了常染色体显性遗传性视网膜色素变性(RP11)剪接因子基因PRPF31中两个突变A194E和A216P的功能后果。利用酵母互补试验，我们证明了将人A216P突变引入酵母同源基因PRP31P中，仅导致在限制性温度下部分恢复生长，表明剪接功能没有完全恢复。使用牛视杆蛋白剪接模板对人体细胞进行的体内剪接功能测试没有发现任何可归因于这两种突变的剪接效率或准确性缺陷，这表明这两种突变对剪接都没有显性的负面影响。然而，对转PRPF31的哺乳动物细胞进行的蛋白质分析和免疫荧光显微镜显示，这两种突变实质上都阻碍了蛋白质移位到细胞核中。因此，总体影响可能是剪接功能不足，只有在剪接需求增加的情况下才会显现出来。随着每天需要补充视盘蛋白，这种情况将存在于视杆感光细胞中，这可能是疾病病理的基础。
This study investigates the functional consequences of two mutations, A194E and A216P, in the splicing factor gene PRPF31 linked to autosomal dominant retinitis pigmentosa (RP11). Using a yeast complementation assay, we demonstrate that introduction of the human A216P mutation into the yeast orthologue PRP31p results in only partial rescue of growth at the restrictive temperature, indicating that splicing function is not fully restored. An in vivo assay of splicing function in human cells using a bovine rod opsin splicing template did not detect any defect in splicing efficiency or accuracy attributable to either mutation, suggesting that neither has a dominant negative effect on splicing. However, western analysis and immunofluorescence microscopy of mammalian cells transfected with PRPF31 revealed that both mutations substantially hinder translocation of the protein into the nucleus. The overall effect may thus be an insufficiency in splicing function, which is revealed only under conditions of elevated splicing demand. With the need to replenish disc proteins on a daily basis, such conditions will exist in rod photoreceptors and this may underlie the disease pathology.