biostudies-literatureChen CWU.S. Department of Health & Human Services | NIH | National Cancer Institute (NCI)Ministry of Science and Technology, TaiwanMinistry of Science and Technology, Taiwan (Ministry of Science and Technology of Taiwan)U.S. Department of Health & Human Services | NIH | National Cancer InstituteNCI NIH HHS2264https://www.ebi.ac.uk/biostudies/studies/S-EPMC10938004biostudies-literatureUnknown15(1)NME3 is a member of the nucleoside diphosphate kinase (NDPK) family localized on the mitochondrial outer membrane (MOM). Here, we report a role of NME3 in hypoxia-induced mitophagy dependent on its active site phosphohistidine but not the NDPK function. Mice carrying a knock-in mutation in the Nme3 gene disrupting NME3 active site histidine phosphorylation are vulnerable to ischemia/reperfusion-induced infarction and develop abnormalities in cerebellar function. Our mechanistic analysis reveals that hypoxia-induced phosphatidic acid (PA) on mitochondria is essential for mitophagy and the interaction of DRP1 with NME3. The PA binding function of MOM-localized NME3 is required for hypoxia-induced mitophagy. Further investigation demonstrates that the interaction with active NME3 prevents DRPNature communicationsNME3 is a gatekeeper for DRP1-dependent mitophagy in hypoxia.PMC10938004NSCT 111-2326-B-002-022CA242443NSCT 111-2634-F-002-017NSCT 111-2320-B-002-088R35 CA242443Hunter TSu CFeng YHsieh STChang ZFCuili XFan CHTing CWYeh TYHuang CYTsai YWChen YJYang KCHuang XRChao TChen CWfalseNME3 is a gatekeeper for DRP1-dependent mitophagy in hypoxia.NME3 is a member of the nucleoside diphosphate kinase (NDPK) family localized on the mitochondrial outer membrane (MOM). Here, we report a role of NME3 in hypoxia-induced mitophagy dependent on its active site phosphohistidine but not the NDPK function. Mice carrying a knock-in mutation in the Nme3 gene disrupting NME3 active site histidine phosphorylation are vulnerable to ischemia/reperfusion-induced infarction and develop abnormalities in cerebellar function. Our mechanistic analysis reveals that hypoxia-induced phosphatidic acid (PA) on mitochondria is essential for mitophagy and the interaction of DRP1 with NME3. The PA binding function of MOM-localized NME3 is required for hypoxia-induced mitophagy. Further investigation demonstrates that the interaction with active NME3 prevents DRP2024-01-01T00:00:00Z2024 Mar2026-07-15T12:01:47.972Z2026-07-04T03:12:28.459ZS-EPMC109380043848068810.1038/s41467-024-46385-7