biostudies-literaturePena IANIDA NIH HHSNHLBI NIH HHSNCI NIH HHSNINDS NIH HHSNIGMS NIH HHS978https://www.ebi.ac.uk/biostudies/studies/S-EPMC11760969biostudies-literatureUnknown16(1)Many essential proteins require pyridoxal 5'-phosphate, the active form of vitamin B6, as a cofactor for their activity. These include enzymes important for amino acid metabolism, one-carbon metabolism, polyamine synthesis, erythropoiesis, and neurotransmitter metabolism. A third of all mammalian pyridoxal 5'-phosphate-dependent enzymes are localized in the mitochondria; however, the molecular machinery involved in the regulation of mitochondrial pyridoxal 5'-phosphate levels in mammals remains unknown. In this study, we used a genome-wide CRISPR interference screen in erythroleukemia cells and organellar metabolomics to identify the mitochondrial inner membrane protein SLC25A38 as a regulator of mitochondrial pyridoxal 5'-phosphate. Loss of SLC25A38 causes depletion of mitochondrial, but Nature communicationsSLC25A38 is required for mitochondrial pyridoxal 5'-phosphate (PLP) accumulation.PMC11760969F31 CA228241F30 CA268633T32 GM007287T32 GM144273T32 HL007118P30 CA014051R35 GM151097T32 GM007753F31 NS127458R00 CA241332P30 DA018343R35 CA242379Yang JVander Heiden MGSienski GAdelmann CHNairn ACLewis CAGe PSabatini DMBlock SBathla SKory NPena IAShi JSKeys HRWitham IHChang SMHeiman MfalseSLC25A38 is required for mitochondrial pyridoxal 5'-phosphate (PLP) accumulation.Many essential proteins require pyridoxal 5'-phosphate, the active form of vitamin B6, as a cofactor for their activity. These include enzymes important for amino acid metabolism, one-carbon metabolism, polyamine synthesis, erythropoiesis, and neurotransmitter metabolism. A third of all mammalian pyridoxal 5'-phosphate-dependent enzymes are localized in the mitochondria; however, the molecular machinery involved in the regulation of mitochondrial pyridoxal 5'-phosphate levels in mammals remains unknown. In this study, we used a genome-wide CRISPR interference screen in erythroleukemia cells and organellar metabolomics to identify the mitochondrial inner membrane protein SLC25A38 as a regulator of mitochondrial pyridoxal 5'-phosphate. Loss of SLC25A38 causes depletion of mitochondrial, but 2025-01-01T00:00:00Z2025 Jan2026-06-02T07:11:15.056Z2025-04-05T00:25:36.274ZS-EPMC117609693985606210.1038/s41467-025-56130-3