Position of premature termination codons determines susceptibility of hERG mutations to nonsense-mediated mRNA decay in long QT syndrome

Qiuming Gong, Matthew R. Stump, Zhengfeng Zhou

    Research output: Research - peer-reviewArticle

    • 7 Citations

    Abstract

    The degradation of human ether-a-go-go-related gene (hERG, KCNH2) transcripts containing premature termination codon (PTC) mutations by nonsense-mediated mRNA decay (NMD) is an important mechanism of long QT syndrome type 2 (LQT2). The mechanisms governing the recognition of PTC-containing hERG transcripts as NMD substrates have not been established. We used a minigene system to study two frameshift mutations, R1032Gfs*25 and D1037Rfs*82. R1032Gfs*25 introduces a PTC in exon 14, whereas D1037Rfs*82 causes a PTC in the last exon (exon 15). We showed that R1032Gfs*25, but not D1037Rfs*82, reduced the level of mutant mRNA compared to the wild-type minigene in an NMD-dependent manner. The deletion of intron 14 prevented degradation of R1032Gfs*25 mRNA indicating that a downstream intron is required for NMD. The recognition and elimination of PTC-containing transcripts by NMD required that the mutation be positioned >. 54-60. nt upstream of the 3'-most exon-exon junction. Finally, we used a full-length hERG splicing-competent construct to show that inhibition of downstream intron splicing by antisense morpholino oligonucleotides inhibited NMD and rescued the functional expression of a third LQT2 mutation, Y1078*. The present study defines the positional requirements for the susceptibility of LQT2 mutations to NMD and posits that the majority of reported LQT2 nonsense and frameshift mutations are potential targets of NMD.

    LanguageEnglish (US)
    Pages190-197
    Number of pages8
    JournalGene
    Volume539
    Issue number2
    DOIs
    StatePublished - Apr 15 2014

    Fingerprint

    Nonsense Mediated mRNA Decay
    Long QT Syndrome
    Nonsense Codon
    Mutation
    Exons
    Introns
    Frameshift Mutation
    Messenger RNA
    Morpholinos
    Antisense Oligonucleotides
    Ether
    Genes

    Keywords

    • Arrhythmia
    • KCNH2
    • Long QT syndrome
    • Patch-clamp
    • Potassium channels

    ASJC Scopus subject areas

    • Genetics

    Cite this

    Position of premature termination codons determines susceptibility of hERG mutations to nonsense-mediated mRNA decay in long QT syndrome. / Gong, Qiuming; Stump, Matthew R.; Zhou, Zhengfeng.

    In: Gene, Vol. 539, No. 2, 15.04.2014, p. 190-197.

    Research output: Research - peer-reviewArticle

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