GAK and PRKCD are positive regulators of PRKN-independent mitophagy

• Published in: Nature communications Vol. 12; no. 1; p. 6101
• Main Authors: Munson, Michael J., Mathai, Benan J., Ng, Matthew Yoke Wui, Trachsel-Moncho, Laura, de la Ballina, Laura R., Schultz, Sebastian W., Aman, Yahyah, Lystad, Alf H., Singh, Sakshi, Singh, Sachin, Wesche, Jørgen, Fang, Evandro F., Simonsen, Anne
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 20.10.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 14/19; 14/35; 14/63; 42/89; 49/1; 631/1647/2163; 631/80/39/2348; 64/11; 64/116; Autophagy; Cyclin G; Damage; Homology; Humanities and Social Sciences; Kinases; Libraries; Lipids; Mitochondria; Mitophagy; multidisciplinary; Phagocytosis; Protein kinase C; Proteins; Regulation; Science; Science (multidisciplinary); siRNA; Zebrafish
• ISSN: 2041-1723; 2041-1723
• 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. The mechanisms involved in programmed or damage-induced removal of mitochondria by mitophagy remain elusive. Here the authors use an siRNA library to screen lipid-binding proteins, and identify the kinases GAK and PRKCD as positive regulators of PRKN-independent mitophagy.