Localization and functional consequences of a direct interaction between TRIOBP-1 and hERG proteins in the heart

David K. Jones, Ashley C. Johnson, Elon C.Roti Roti, Fang Liu, Rebecca Uelmen, Rebecca A. Ayers, Istvan Baczko, David J. Tester, Michael J. Ackerman, Matthew C. Trudeau, Gail A. Robertson

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Reduced levels of the cardiac human (h)ERG ion channel protein and the corresponding repolarizing current IKr can cause arrhythmia and sudden cardiac death, but the underlying cellular mechanisms controlling hERG surface expression are not well understood. Here, we identified TRIOBP-1, an F-actin-binding protein previously associated with actin polymerization, as a putative hERGinteracting protein in a yeast-two hybrid screen of a cardiac library. We corroborated this interaction by performing Förster resonance energy transfer (FRET) in HEK293 cells and co-immunoprecipitation in HEK293 cells and native cardiac tissue. TRIOBP-1 overexpression reduced hERG surface expression and current density, whereas reducing TRIOBP-1 expression via shRNA knockdown resulted in increased hERG protein levels. Immunolabeling in rat cardiomyocytes showed that native TRIOBP-1 colocalized predominantly with myosin-binding protein C and secondarily with rat ERG. In human stem cell-derived cardiomyocytes, TRIOBP-1 overexpression caused intracellular co-sequestration of hERG signal, reduced native IKr and disrupted action potential repolarization. Ca2+ currents were also somewhat reduced and cell capacitance was increased. These findings establish that TRIOBP-1 interacts directly with hERG and can affect protein levels, IKr magnitude and cardiac membrane excitability.

Original languageEnglish
Article numberjcs206730
JournalJournal of cell science
Volume131
Issue number6
DOIs
Publication statusPublished - Mar 1 2018

Keywords

  • Action potential
  • FRET
  • I
  • IPSC-CM
  • KCNH2
  • TARA
  • Yeast two-hybrid

ASJC Scopus subject areas

  • Cell Biology

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    Jones, D. K., Johnson, A. C., Roti, E. C. R., Liu, F., Uelmen, R., Ayers, R. A., Baczko, I., Tester, D. J., Ackerman, M. J., Trudeau, M. C., & Robertson, G. A. (2018). Localization and functional consequences of a direct interaction between TRIOBP-1 and hERG proteins in the heart. Journal of cell science, 131(6), [jcs206730]. https://doi.org/10.1242/jcs.206730