Division of Biochemistry, Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, N-0372, Oslo, Norway.
Centre for Cancer Cell Reprogramming, Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, N-0316, Oslo, Norway.
Nat Commun. 2021 Oct 20;12(1):6101. doi: 10.1038/s41467-021-26331-7.
The mechanisms involved in programmed or damage-induced removal of mitochondria by mitophagy remains elusive. Here, we have screened for regulators of PRKN-independent mitophagy using an siRNA library targeting 197 proteins containing lipid interacting domains. We identify Cyclin G-associated kinase (GAK) and Protein Kinase C Delta (PRKCD) as regulators of PRKN-independent mitophagy, with both being dispensable for PRKN-dependent mitophagy and starvation-induced autophagy. We demonstrate that the kinase activity of both GAK and PRKCD are required for efficient mitophagy in vitro, that PRKCD is present on mitochondria, and that PRKCD facilitates recruitment of ULK1/ATG13 to early autophagic structures. Importantly, we demonstrate in vivo relevance for both kinases in the regulation of basal mitophagy. Knockdown of GAK homologue (gakh-1) in C. elegans or knockout of PRKCD homologues in zebrafish led to significant inhibition of basal mitophagy, highlighting the evolutionary relevance of these kinases in mitophagy regulation.
线粒体通过自噬作用程序性或损伤诱导清除的机制仍然难以捉摸。在这里，我们使用靶向含有脂质相互作用域的 197 种蛋白质的 siRNA 文库筛选了 PRKN 非依赖性自噬的调节剂。我们确定细胞周期蛋白 G 相关激酶 (GAK) 和蛋白激酶 C Delta (PRKCD) 是 PRKN 非依赖性自噬的调节剂，两者对于 PRKN 依赖性自噬和饥饿诱导的自噬都是可有可无的。我们证明了 GAK 和 PRKCD 的激酶活性都需要体外有效的自噬作用，PRKCD 存在于线粒体上，并且 PRKCD 促进 ULK1/ATG13 向早期自噬结构的募集。重要的是，我们体内证明了这两种激酶在调节基础自噬中的相关性。在秀丽隐杆线虫中敲低 GAK 同源物 (gakh-1) 或在斑马鱼中敲除 PRKCD 同源物导致基础自噬显著抑制，突出了这些激酶在自噬调节中的进化相关性。
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