Pyrimidines maintain mitochondrial pyruvate oxidation to support de novo lipogenesis

This study elucidates the essential role of pyrimidines in supporting mitochondrial pyruvate oxidation and the tricarboxylic acid (TCA) cycle. Quantitative assessment of metabolite changes in response to alterations in cellular pyrimidine levels was conducted using liquid chromatography-mass spectrometry. Concurrently, western blot analysis was employed to characterize the expression of metabolic enzymes potentially involved in this regulatory process. To enhance precision, metabolic activity measurements were conducted under both untreated and pyrimidine-depleted conditions using radioactivity-based assays, isotope tracers, and Seahorse analyzers. The comprehensive dataset includes experimental quantifications of metabolite abundance, accurate assessments of metabolic activity, and protein levels of metabolic and signaling enzymes.