be230cLecture9

Rt pcr analysis revealed that the 23981gc mutation

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Unformatted text preview: site in intron 9, leading to the inclusion of 54 nt of the intron 9 sequence in hERG mRNA. The cryptic splicing resulted in an in-frame insertion of Abstract 18 amino acids in the middle of the cyclic nucleotide binding domain. In patch clamp experiments Mutations in the human ether-a-go-go-related gene (hERG) cause type 2 long QT syndrome. In this the splice mutant did not generate hERG current. Western blot and immunostaining studies showed study, we investigated the mechanism of the hERG splice site mutation 2398+1G>C and the that the mutant expressed an immature form of hERG protein that failed to reach the plasma genotype-phenotype relationship of mutation carriers inwild-type channels led to a with long negative three unrelated kindreds dominant QT membrane. Coexpression of the mutant and syndrome. The effect of 2398+1G>C channel function by intracellular retention of heteromeric channels. Our suppression of wild-type on mRNA splicing was studied by analysis of RNA isolated from lymphocytes of index patients 2398+1G>C activates a cryptic site and generates a full-length hERG protein results demonstrate that and using minigenes expressed in HEK293 cells and neonatal rat ventricular myocytes. RT-PCR analysis revealed that theto a traffickingmutation disrupted the with an insertion of 18 amino acids, which leads 2398+1G>C defect of the mutant channel. NIH-PA Author Man normal splicing and activated a cryptic splice donor site in intron 9, leading to the inclusion of 54 nt of the intron 9 sequence in hERG mRNA. The cryptic splicing resulted in an in-frame insertion of 18 aminoKeywords middle of the cyclic nucleotide binding domain. In patch clamp experiments acids in the the splice mutant did not generate hERG mutation;Western blotsudden death; myocytes long QT syndrome; splicing current. arrhythmia; and immunostaining studies showed that the mutant expressed an immature form of hERG protein that failed to reach the plasma membrane. Coexpression of the mutant and wild-type channels led to a dominant negative suppression Introduction 1. of w...
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