Sequence and structure-specific elements of HERG mRNA determine channel synthesis and trafficking efficiency

Jakub Sroubek, Yamini Krishnan, Thomas V. McDonald

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Human ether-á-gogo-related gene (HERG) encodes a potassium channel that is highly susceptible to deleterious mutations resulting in susceptibility to fatal cardiac arrhythmias. Most mutations adversely affect HERG channel assembly and trafficking. Why the channel is so vulnerable to missense mutations is not well understood. Since nothing is known of how mRNA structural elements factor in channel processing, we synthesized a codon-modified HERG cDNA (HERG-CM) where the codons were synonymously changed to reduce GC content, secondary structure, and rare codon usage. HERG-CM produced typical IKr-like currents; however, channel synthesis and processing were markedly different. Translation efficiency was reduced for HERG-CM, as determined by heterologous expression, in vitro translation, and polysomal profiling. Trafficking efficiency to the cell surface was greatly enhanced, as assayed by immunofluorescence, subcellular fractionation, and surface labeling. Chimeras of HERG-NT/CM indicated that trafficking efficiency was largely dependent on 5= sequences, while translation efficiency involved multiple areas. These results suggest that HERG translation and trafficking rates are independently governed by noncoding information in various regions of the mRNA molecule. Noncoding information embedded within the mRNA may play a role in the pathogenesis of hereditary arrhythmia syndromes and could provide an avenue for targeted therapeutics.

Original languageEnglish (US)
Pages (from-to)3039-3053
Number of pages15
JournalFASEB Journal
Volume27
Issue number8
DOIs
StatePublished - Aug 2013

Fingerprint

Ether
Genes
Messenger RNA
Codon
Cardiac Arrhythmias
Mutation
Potassium Channels
Base Composition
Missense Mutation
Fractionation
Processing
Labeling
Fluorescent Antibody Technique
Complementary DNA
Molecules

Keywords

  • Codon-usage
  • KCNH
  • LQTS
  • Protein translation
  • SNP

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology
  • Medicine(all)

Cite this

Sequence and structure-specific elements of HERG mRNA determine channel synthesis and trafficking efficiency. / Sroubek, Jakub; Krishnan, Yamini; McDonald, Thomas V.

In: FASEB Journal, Vol. 27, No. 8, 08.2013, p. 3039-3053.

Research output: Contribution to journalArticle

Sroubek, Jakub ; Krishnan, Yamini ; McDonald, Thomas V. / Sequence and structure-specific elements of HERG mRNA determine channel synthesis and trafficking efficiency. In: FASEB Journal. 2013 ; Vol. 27, No. 8. pp. 3039-3053.
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