Multiple splicing defects caused by hERG splice site mutation 2592+1G>A associated with long QT syndrome.
Multiple splicing defects caused by hERG splice site mutation 2592+1G>A associated with long QT syndrome.
复制标题
hERG 剪接位点突变 2592 1G>A 引起的多重剪接缺陷与长 QT 综合征相关。
DOI：
10.1152/ajpheart.00818.2010
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Zhou,Zhengfeng
中科院分区：
文献类型：
--
作者：
Stump,MatthewR;Gong,Qiuming;Zhou,Zhengfeng
Long QT syndrome type 2 (LQT2) is caused by mutations in the human ether-a-go-go-related gene (hERG). Cryptic splice site activation inhERGhas recently been identified as a novel pathogenic mechanism of LQT2. In this report, we characterize ahERGsplice site mutation, 2592+1G>A, which occurs at the 5′ splice site of intron 10. Reverse transcription-PCR analyses using hERG minigenes transfected into human embryonic kidney-293 cells and HL-1 cardiomyocytes revealed that the 2592+1G>A mutation disrupted normal splicing and caused multiple splicing defects: the activation of cryptic splice sites within exon 10 and intron 10 and complete intron 10 retention. We performed functional and biochemical analyses of the major splice product, hERGΔ24, in which 24 amino acids within the cyclic nucleotide binding domain of the hERG channel COOH-terminus is deleted. Patch-clamp experiments revealed that the splice mutant did not generate hERG current. Western blot and immunostaining studies showed that mutant channels did not traffic to the cell surface. Coexpression of wild-type hERG and hERGΔ24 resulted in significant dominant-negative suppression of hERG current via the intracellular retention of the wild-type channels. Our results demonstrate that 2592+1G>A causes multiple splicing defects, consistent with the pathogenic mechanisms of long QT syndrome.
