We found that UBE3A regulates the stability and enzymaticactivity of PDHA1 and ACAT1 in the cell, and the overexpression ofUBE3A in the mouse liver would enhance glycolysis and ketogenesisto condition the liver for lipid accumulation as a primer for NAFLD.Our findings unveiled a new axis enabling the regulation of glucoseand lipid metabolism by protein ubiquitination and suggested a potentialrole for UBE3A in NAFLD development. Here, UBE3A is linked to metabolic dysfunction-associated steatotic liver disease.