Fatty acid metabolism controls plasma membrane cholesterol accessibility via ATGL-dependent phospholipid remodeling

Accessible cholesterol, the pool of membrane cholesterol with high chemical activity, regulates vital processes including vertebrate development and pathogen evasion. The mechanisms that govern plasma membrane (PM) cholesterol accessibility are incompletely understood. Using a genome-wide screen we find that acetyl-CoA carboxylase alpha (ACC1) loss causes a ~10-fold increase in PM accessible cholesterol in cells and a male mouse model. We demonstrate that reduced fatty acyl-CoA levels, achieved by targeting metabolic enzymes including ACC1, fatty acid synthase (FASN) or acyl-CoA synthetases long chain (ACSL1, 3 and 4), results in the activation of adipose triacylglycerol lipase (ATGL). ATGL activation elevates polyunsaturated diacylglycerols even in triacylglycerol-deficient backgrounds, suggesting that ATGL acts independently of triacylglycerol hydrolysis. These diacylglycerol species are utilized to generate polyunsaturated phosphatidylcholine and phosphatidylethanolamine, which raises PM fluidity and cholesterol accessibility. Conversely, ATGL inhibition rigidifies the PM, reducing PM accessible cholesterol. Increased PM fluidity impairs cholesterol transport, triggering SREBP2 activation. This study reveals a surprising link between fatty acid metabolism and cholesterol homeostasis, demonstrating how ATGL-dependent lipid remodeling tunes membrane fluidity to regulate intracellular cholesterol signaling.