ABC transporters control ATP release through cholesterol-dependent volume-regulated anion channel activity
ABC transporters control ATP release through cholesterol-dependent volume-regulated anion channel activity
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DOI：
10.1074/jbc.ra119.010699
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发表时间：
2020-04-17
影响因子：
4.8
通讯作者：
Tomita, Susumu
中科院分区：
文献类型：
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作者：
Dunn, Patrick J.;Salm, Elizabeth J.;Tomita, Susumu
嘌呤能信号通过细胞外ATP调节多种细胞事件，并涉及正常生理和病理生理。一些分子与ATP和其他小分子的释放有关，但由于它们的复杂性和异质性，它们的精确贡献很难评估。在这里，我们报告了使用HEK-293细胞和小鼠小脑颗粒神经元，以及生物发光、钙flir和短发卡RNA，对低张力诱导的ATP释放调节剂的功能获得筛选结果。基于基因沉默分析。该筛选利用了迄今为止最广泛的全基因组ORF收集，覆盖了90%的人类非冗余蛋白质编码基因。我们发现了两个ABCG1 （ABC亚家族G成员1）变体，它们调节细胞胆固醇，作为低张力诱导的ATP释放的调节剂。我们发现胆固醇水平控制着容量调节的阴离子通道？依赖性ATP释放。这些发现揭示了ATP释放和负离子通道活性调节的新机制，并提供了细胞状态、胆固醇和嘌呤能信号传导之间的关键联系。
Purinergic signaling by extracellular ATP regulates a variety of cellular events and is implicated in both normal physiology and pathophysiology. Several molecules have been associated with the release of ATP and other small molecules, but their precise contributions have been difficult to assess because of their complexity and heterogeneity. Here, we report on the results of a gain-of-function screen for modulators of hypotonicity-induced ATP release using HEK-293 cells and murine cerebellar granule neurons, along with bioluminescence, calcium FLIPR, and short hairpin RNA?based gene-silencing assays. This screen utilized the most extensive genome-wide ORF collection to date, covering 90% of human, nonredundant, protein-encoding genes. We identified two ABCG1 (ABC subfamily G member 1) variants, which regulate cellular cholesterol, as modulators of hypotonicity-induced ATP release. We found that cholesterol levels control volume-regulated anion channel?dependent ATP release. These findings reveal novel mechanisms for the regulation of ATP release and volume-regulated anion channel activity and provide critical links among cellular status, cholesterol, and purinergic signaling.