Cardiac Nav1.5 is modulated by ubiquitin protein ligase E3 component n‐recognin UBR3 and 6

Journal of Cellular and Molecular Medicine

Chunxia Zhao¹  Lijie Wang¹  Xiue Ma¹  Weidong Zhu¹  Lei Yao¹  Yingyu Cui¹  Yi Liu¹  Jun Li¹  Xingqun Liang¹  Yunfu Sun¹  Li Li¹ 

[1] Key Laboratory of Arrhythmias of the Ministry of Education of China, East Hospital, Tongji University School of Medicine, Shanghai, China

关键词: Nav1.5 channel;    UBR;    cardiomyocyte;    degradation;    ubiquitin;    proteasome;   

DOI  :  10.1111/jcmm.12588

来源: Wiley

【 摘 要 】

Abstract

The voltage-gated Na⁺ channel Na_v1.5 is essential for action potential (AP) formation and electrophysiological homoeostasis in the heart. The ubiquitin–proteasome system (UPS) is a major degradative system for intracellular proteins including ion channels. The ubiquitin protein ligase E3 component N-recognin (UBR) family is a part of the UPS; however, their roles in regulating cardiac Na_v1.5 channels remain elusive. Here, we found that all of the UBR members were expressed in cardiomyocytes. Individual knockdown of UBR3 or UBR6, but not of other UBR members, significantly increased Na_v1.5 protein levels in neonatal rat ventricular myocytes, and this effect was verified in HEK293T cells expressing Na_v1.5 channels. The UBR3/6-dependent regulation of Na_v1.5 channels was not transcriptionally mediated, and pharmacological inhibition of protein biosynthesis failed to counteract the increase in Na_v1.5 protein caused by UBR3/6 reduction, suggesting a degradative modulation of UBR3/6 on Na_v1.5. Furthermore, the effects of UBR3/6 knockdown on Na_v1.5 proteins were abolished under the inhibition of proteasome activity, and UBR3/6 knockdown reduced Na_v1.5 ubiquitylation. The double UBR3–UBR6 knockdown resulted in comparable increases in Na_v1.5 proteins to that observed for single knockdown of either UBR3 or UBR6. Electrophysiological recordings showed that UBR3/6 reduction-mediated increase in Na_v1.5 protein enhanced the opening of Na_v1.5 channels and thereby the amplitude of the AP. Thus, our findings indicate that UBR3/6 regulate cardiomyocyte Na_v1.5 channel protein levels via the ubiquitin–proteasome pathway. It is likely that UBR3/6 have the potential to be a therapeutic target for cardiac arrhythmias.

【 授权许可】

CC BY   
© 2015 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

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