title: Alternative promoter usage and splicing of the human SCN5A gene contribute to transcript heterogeneity.
abstract: The sodium channel isoform Na(v)1.5 mediates sodium current, excitability, and electrical conduction in the human heart. Recent studies have indicated alternative splicing within the protein-coding portion of its gene, SCN5A, as a mechanism to generate diversity in Na(v)1.5 protein structure and function. In the present study we identified several novel SCN5A transcripts in human heart, displaying distinct 5′-untranslated regions but identical protein-coding sequences. These transcripts originated from the splicing of alternative exons 1 (designated 1A, 1B, 1C, and 1D) to the translational start codon-containing exon 2, and were preferentially expressed in the heart as compared to other tissues. Comparison of their expression level between adult and fetal heart demonstrated that exon 1C- and 1D-derived sequences were more prominent in adult than in fetal heart. Two new promoters (designated P2 and P3) for the SCN5A gene were identified and functionally characterized in myocardial- and nonmyocardial-derived cell lines. Translation of the transcript containing exon 1D-derived sequences proved to be significantly impaired in these cell lines, which could be restored by mutation of an upstream translational start codon. These results implicate the usage of alternative promoters and 5′-untranslated regions as new mechanisms in the regulation of human Na(v)1.5 expression.
authors.affiliation: Division of Heart and Lungs, Department of Medical Physiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Leonie
authors.initials: L
authors.lastname: van Stuijvenberg
authors.forename: Cansu
authors.initials: C
authors.lastname: Yildirim
authors.forename: Bart G J M
authors.initials: BG
authors.lastname: Kok
authors.forename: Toon A B
authors.initials: TA
authors.lastname: van Veen
authors.forename: András
authors.initials: A
authors.lastname: Varró
authors.forename: Stephan K G
authors.initials: SK
authors.lastname: Winckels
authors.forename: Marc A
authors.initials: MA
authors.lastname: Vos
authors.forename: Marti F A
authors.initials: MF
authors.lastname: Bierhuizen
mesh_terms: D000328:Adult; D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002460:Cell Line; D002478:Cells, Cultured; D018387:Codon, Initiator / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D006321:Heart / Q000196:embryology; D006801:Humans; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000235:genetics* / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D062554:NAV1.5 Voltage-Gated Sodium Channel; D016366:Open Reading Frames; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing; D051381:Rats; D015222:Sodium Channels / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D018387:Codon, Initiator; D009124:Muscle Proteins; D062554:NAV1.5 Voltage-Gated Sodium Channel; D020033:Protein Isoforms; C568320:SCN5A protein, human; C568322:Scn5a protein, rat; D015222:Sodium Channels