UC Davis

Background

Long QT syndrome (LQTS) is a hereditary disease that predisposes patients to life-threatening cardiac arrhythmias and sudden cardiac death. Our previous study of the human ether-à-go-go related gene (hERG)-encoded K⁺ channel (K_v11.1) supports an association between hERG and RING finger protein 207 (RNF207) variants in aggravating the onset and severity of LQTS, specifically T613M hERG (hERG_T613M) and RNF207 frameshift (RNF207_G603fs) mutations. However, the underlying mechanistic underpinning remains unknown.

Objective

The purpose of the present study was to test the role of RNF207 in the function of hERG-encoded K⁺ channel subunits.

Methods

Whole-cell patch-clamp experiments were performed in human embryonic kidney (HEK 293) cells and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) together with immunofluorescent confocal and high resolution microscopy, auto-ubiquitinylation assays, and co-immunoprecipitation experiments to test the functional interactions between hERG and RNF207.

Results

Here, we demonstrated that RNF207 serves as an E3 ubiquitin ligase and targets misfolded hERG_T613M proteins for degradation. RNF207_G603fs exhibits decreased activity and hinders the normal degradation pathway; this increases the levels of hERG_T613M subunits and their dominant-negative effect on the wild-type subunits, ultimately resulting in decreased current density. Similar findings are shown for hERG_A614V, a known dominant-negative mutant subunit. Finally, the presence of RNF207_G603fs with hERG_T613M results in significantly prolonged action potential durations and reduced hERG current in human-induced pluripotent stem cell-derived cardiomyocytes.

Conclusion

Our study establishes RNF207 as an interacting protein serving as a ubiquitin ligase for hERG-encoded K⁺ channel subunits. Normal function of RNF207 is critical for the quality control of hERG subunits and consequently cardiac repolarization. Moreover, our study provides evidence for protein quality control as a new paradigm in life-threatening cardiac arrhythmias in patients with LQTS.