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 John 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 (US)
Article numberjcs206730
JournalJournal of Cell Science
Volume131
Issue number6
DOIs
StatePublished - Mar 1 2018

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HEK293 Cells
Cardiac Myocytes
Proteins
Energy Transfer
Sudden Cardiac Death
Ion Channels
Immunoprecipitation
Polymerization
Small Interfering RNA
Action Potentials
Libraries
Actins
Cardiac Arrhythmias
Stem Cells
Yeasts
Membranes
F-actin-binding proteins
myosin-binding protein C

Keywords

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

ASJC Scopus subject areas

  • Cell Biology

Cite this

Jones, D. K., Johnson, A. C., Roti, E. C. R., Liu, F., Uelmen, R., Ayers, R. A., ... 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

Localization and functional consequences of a direct interaction between TRIOBP-1 and hERG proteins in the heart. / Jones, David K.; Johnson, Ashley C.; Roti, Elon C.Roti; Liu, Fang; Uelmen, Rebecca; Ayers, Rebecca A.; Baczko, Istvan; Tester, David J.; Ackerman, Michael John; Trudeau, Matthew C.; Robertson, Gail A.

In: Journal of Cell Science, Vol. 131, No. 6, jcs206730, 01.03.2018.

Research output: Contribution to journalArticle

Jones, DK, Johnson, AC, Roti, ECR, Liu, F, Uelmen, R, Ayers, RA, Baczko, I, Tester, DJ, Ackerman, MJ, Trudeau, MC & Robertson, GA 2018, 'Localization and functional consequences of a direct interaction between TRIOBP-1 and hERG proteins in the heart', Journal of Cell Science, vol. 131, no. 6, jcs206730. https://doi.org/10.1242/jcs.206730
Jones, David K. ; Johnson, Ashley C. ; Roti, Elon C.Roti ; Liu, Fang ; Uelmen, Rebecca ; Ayers, Rebecca A. ; Baczko, Istvan ; Tester, David J. ; Ackerman, Michael John ; Trudeau, Matthew C. ; Robertson, Gail A. / Localization and functional consequences of a direct interaction between TRIOBP-1 and hERG proteins in the heart. In: Journal of Cell Science. 2018 ; Vol. 131, No. 6.
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