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UBC9 regulates cardiac sodium channel Na v 1.5 ubiquitination, degradation and sodium current density.
- Source :
-
Journal of molecular and cellular cardiology [J Mol Cell Cardiol] 2019 Apr; Vol. 129, pp. 79-91. Date of Electronic Publication: 2019 Feb 14. - Publication Year :
- 2019
-
Abstract
- Voltage-gated sodium channel Na <subscript>v</subscript> 1.5 is critical for generation and conduction of cardiac action potentials. Mutations and expression level changes of Na <subscript>v</subscript> 1.5 are associated with cardiac arrhythmias and sudden death. The ubiquitin (Ub) conjugation machinery utilizes three enzyme activities, E1, E2, and E3, to regulate protein degradation. Previous studies from us and others showed that Nedd4-2 acts as an E3 ubiquitin-protein ligase involved in ubiquitination and degradation of Na <subscript>v</subscript> 1.5, however, more key regulators remain to be identified. In this study, we show that UBC9, a SUMO-conjugating enzyme, regulates ubiquitination and degradation of Na <subscript>v</subscript> 1.5. Overexpression of UBC9 significantly decreased Na <subscript>v</subscript> 1.5 expression and reduced sodium current densities, whereas knockdown of UBC9 expression significantly enhanced Na <subscript>v</subscript> 1.5 expression and increased sodium current densities, in both HEK293 cells and primary neonatal cardiomyocytes. Overexpression of UBC9 increased ubiquitination of Na <subscript>v</subscript> 1.5, and proteasome inhibitor MG132 blocked the effect of UBC9 overexpression on Na <subscript>v</subscript> 1.5 degradation. Co-immunoprecipitation showed that UBC9 interacts with Nedd4-2. UBC9 with mutation C93S, which suppresses SUMO-conjugating activity of UBC9, was as active as wild type UBC9 in regulating Na <subscript>v</subscript> 1.5 levels, suggesting that UBC9 regulates Na <subscript>v</subscript> 1.5 expression levels in a SUMOylation-independent manner. Our findings thus identify a key structural element of the ubiquitin-conjugation machinery for Na <subscript>v</subscript> 1.5 and provide important insights into the regulatory mechanism for ubiquitination and turnover of Na <subscript>v</subscript> 1.5.<br /> (Copyright © 2019 Elsevier Ltd. All rights reserved.)
- Subjects :
- Animals
Animals, Newborn
Down-Regulation genetics
HEK293 Cells
HeLa Cells
Humans
Nedd4 Ubiquitin Protein Ligases metabolism
Proteasome Endopeptidase Complex metabolism
Protein Binding
Rats
Small Ubiquitin-Related Modifier Proteins metabolism
Sumoylation
Ubiquitin-Conjugating Enzymes genetics
Up-Regulation genetics
Ion Channel Gating
NAV1.5 Voltage-Gated Sodium Channel metabolism
Proteolysis
Sodium metabolism
Ubiquitin-Conjugating Enzymes metabolism
Ubiquitination
Subjects
Details
- Language :
- English
- ISSN :
- 1095-8584
- Volume :
- 129
- Database :
- MEDLINE
- Journal :
- Journal of molecular and cellular cardiology
- Publication Type :
- Academic Journal
- Accession number :
- 30772377
- Full Text :
- https://doi.org/10.1016/j.yjmcc.2019.02.007