Synergy between RNA editing and alternative splicing modulates the biological properties of the voltage-gated calcium channel CaV1.3

Abstract

Alternative splicing and adenosine-to-inosine (A-to-I) RNA editing are post-transcriptional mechanisms that promote proteomic diversity in the brain. The impact of RNA editing on brain physiology is underscored by its association with the etiology and pathogenesis of various neurological and neurodegenerative diseases. Alternative splicing of exon 43 within the CaV1.3 C-terminus produces long (CaV1.3L) and short (CaV1.343S) variants exhibiting distinct gating and pharmacological properties. While the physiological role of CaV1.3 A-to-I RNA editing in brain function is still unfolding, recent findings indicate that unedited CaV1.3 channels display larger calcium (Ca2+) influx and enhanced neuronal excitability, suggesting that A-to-I RNA editing may act as a negative regulator of neuronal Ca2+ signaling. However, it remains unclear which specific splice variants of CaV1.3 undergo RNA editing. We discovered an unexpected link between alternative splicing and RNA editing, where generation of the CaV1.343S variant results in a 3-fold increase in RNA editing at the IQ site. The CaV1.343S variant undergoes A-to-I RNA editing at 2 sites within its calmodulin-binding IQ domain, giving rise to 3 different edited variants. Functional analysis revealed that A-to-I RNA editing markedly decreases current density and induces a depolarizing shift in the current window, aligning the edited short variant's properties with those of the long variant. Our findings suggest that A-to-I RNA editing serves as a physiological mechanism regulating the 'short' gating properties of the CaV1.343S variant and positions A-to-I RNA editing of CaV1.343S variant as a neuroprotective physiological mechanism that prevents Ca2+ overload, especially in neurons susceptible to Ca2+ toxicity.