O-GlcNAcylation of fatty acid synthase is required for its proper subcellular localization, expression level, and activity.

Fatty Acid Synthase (FASN) is involved in various fundamental cellular processes through its pivotal role in producing fatty acids through the de novo lipogenesis pathway. FASN is frequently overexpressed in tumors and participates in cancer cell proliferation. Little has been documented regarding post-translational modifications of FASN. We previously demonstrated that O-GlcNAcylation regulates FASN in mice livers and in the HepG2 hepatic cancer cell line. In the present study, we show that modulation of global O-GlcNAcylation levels impacts fatty acids production in HepG2 cells. We identified serine 595 and threonine 980 as major O-GlcNAcylation sites. While mutation of S595 moderately affects FASN behavior, T980 is crucial for FASN expression, membrane localization, homodimerization, stability, and activity in Hep3B cells. This residue is necessary for FASN properties, promoting cell survival, cell proliferation, and cell cycle progression. Our results suggest that targeting FASN at T980 may open an interesting path for controlling its catalytic activity.