Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan, USA.
Department of Molecular & Integrative Physiology, University of Michigan Medical School, Ann Arbor, Michigan, USA.
JCI Insight. 2021 Feb 8;6(3):141776. doi: 10.1172/jci.insight.141776.
Loss-of-function (LOF) variants in SCN1B, encoding voltage-gated sodium channel β1 subunits, are linked to human diseases with high risk of sudden death, including developmental and epileptic encephalopathy and cardiac arrhythmia. β1 Subunits modulate the cell-surface localization, gating, and kinetics of sodium channel pore-forming α subunits. They also participate in cell-cell and cell-matrix adhesion, resulting in intracellular signal transduction, promotion of cell migration, calcium handling, and regulation of cell morphology. Here, we investigated regulated intramembrane proteolysis (RIP) of β1 by BACE1 and γ-secretase and show that β1 subunits are substrates for sequential RIP by BACE1 and γ-secretase, resulting in the generation of a soluble intracellular domain (ICD) that is translocated to the nucleus. Using RNA sequencing, we identified a subset of genes that are downregulated by β1-ICD overexpression in heterologous cells but upregulated in Scn1b-null cardiac tissue, which lacks β1-ICD signaling, suggesting that the β1-ICD may normally function as a molecular brake on gene transcription in vivo. We propose that human disease variants resulting in SCN1B LOF cause transcriptional dysregulation that contributes to altered excitability. Moreover, these results provide important insights into the mechanism of SCN1B-linked channelopathies, adding RIP-excitation coupling to the multifunctionality of sodium channel β1 subunits.
失活(LOF)变体在 SCN1B 中,编码电压门控钠离子通道β1 亚基,与具有高猝死风险的人类疾病有关,包括发育性和癫痫性脑病和心律失常。β1 亚基调节钠通道孔形成α亚基的细胞表面定位、门控和动力学。它们还参与细胞-细胞和细胞-基质黏附,从而导致细胞内信号转导、促进细胞迁移、钙处理和细胞形态调节。在这里,我们研究了 BACE1 和 γ-分泌酶对β1 的调节跨膜蛋白水解(RIP),并表明β1 亚基是 BACE1 和 γ-分泌酶顺序 RIP 的底物,导致可溶细胞内结构域(ICD)的产生,该结构域易位到细胞核。使用 RNA 测序,我们鉴定了一组基因,这些基因在异源细胞中β1-ICD 过表达时下调,但在缺乏β1-ICD 信号的 Scn1b 缺失心脏组织中上调,这表明β1-ICD 可能在体内通常作为转录的分子制动器。我们提出,导致 SCN1B LOF 的人类疾病变体引起转录失调,导致兴奋性改变。此外,这些结果为 SCN1B 相关通道病的机制提供了重要的见解,将 RIP-兴奋偶联添加到钠通道β1 亚基的多功能性中。