PRRT2 truncated mutations lead to nonsense-mediated mRNA decay in Paroxysmal Kinesigenic Dyskinesia.
PRRT2 truncated mutations lead to nonsense-mediated mRNA decay in Paroxysmal Kinesigenic Dyskinesia.
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DOI：
10.1016/j.parkreldis.2014.10.012
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发表时间：
2014-12
影响因子：
4.1
通讯作者：
Li Wu;Huidong Tang;Xiao-jun Huang;Lan Zheng;Xiao-li Liu;Tian Wang;Jing-yi Wang;Li Cao;S. Chen
中科院分区：
文献类型：
--
作者：
Li Wu;Huidong Tang;Xiao-jun Huang;Lan Zheng;Xiao-li Liu;Tian Wang;Jing-yi Wang;Li Cao;S. Chen
背景与目的阵发性运动诱发性运动障碍（Paroxysmal Kinesigenic Dyskinesia，PKD）是一种以反复发作和短暂的不自主运动为特征的发作性不自主运动障碍。富含脯氨酸的跨膜蛋白2（PRRT 2）已被确定为PKD、良性家族性婴儿惊厥（BFIC）和伴有舞蹈手足徐动症的婴儿惊厥（ICCA）的致病基因。此外，在PED或PNKD患者中也检测到PRRT 2突变。方法我们用无义介导的mRNA衰变（NMD）途径的抑制剂--盐酸乙双胍和放线菌酮以及无义转录物沉默调节因子1（UPF 1）与永生化淋巴母细胞一起检测截短突变是否导致NMD，到目前为止，在每个真核细胞中证明了一种mRNA监视类型，并且通常降解含有过早翻译终止密码子（PTC）的mRNA。此外，我们用野生型和突变型PRRT 2质粒转染SH-SY 5 Y细胞，以确定PRRT 2蛋白的亚细胞localization.ResultsWe检测到，截短的PRRT 2的低表达，并通过应用NMD通路的抑制剂进一步获救，这表明NMD在PKD的发病机制中起着重要作用的单倍不足。此外，对于一小部分未降解的突变体PRRT 2被翻译成截短的蛋白质，其细胞定位从膜到细胞质和核，这可能会导致一个功能loss.ConclusionWe建议的NMD的PRRT 2的截短突变和改变细胞定位的未降解的PRRT 2，可能会导致PKD。
Background and purposeParoxysmal Kinesigenic Dyskinesia (PKD) is an episodic involuntary movement disorder characterized by recurrent and brief involuntary movements. Proline-rich transmembrane protein 2 (PRRT2) has been identified as the causative gene for PKD, Benign familial infantile convulsions (BFIC) and Infantile convulsions with choreoathetosis (ICCA). As well, PRRT2 mutations have been detected in patients with PED or PNKD. To date, most of the mutations have been found to be nonsense.MethodWe used inhibitors of nonsense-mediated mRNA decay (NMD) pathway --emetine dihydrochloride hydrate and cycloheximide and silencing regulator of nonsense transcripts 1(UPF1) with immortalized lymphoblasts to detect whether the truncated mutations lead to NMD, a type of mRNA surveillance in every eukaryotic cell proved so far and that generally degrades mRNA containing premature translation termination codons (PTCs). In addition, we transfected the SH-SY5Y cells with wild-type and mutant PRRT2 plasmids to identify the PRRT2 protein's subcellular localization.ResultsWe detected, low expression of truncated PRRT2 and was further rescued by applying the inhibitor of NMD pathway, suggesting that NMD plays an important role in the pathogenesis of PKD by haplo-insufficiency. Moreover, for the small portion of undegraded mutant PRRT2 that was translated into truncated proteins, their cellular localization changed from membrane to cytoplasm and nuclear, which might lead to a functional loss.ConclusionWe suggest that the NMD of truncated mutation of PRRT2 and altered cellular localization of undegraded of PRRT2, might lead to PKD.