STIMULATION OF ACYL-COA-CHOLESTEROL ACYLTRANSFERASE ACTIVITY BY BREFELDIN-A IN MACROPHAGE J774 CELLS

Brefeldin A (BFA), an inhibitor of secretory pathway, enhances incorporation of radiolabeled cholesterol and oleate into cholesteryl esters in cultured cells [12]. We studied the mechanism for this effect of BFA in the macrophage J774. When incubated with 2.7 muM BFA in the absence of lipoproteins, J774 cells synthesized and accumulated 1.5- to 4-fold more cholesteryl esters than did cells which received no BFA. BFA caused neither an elevation of cholesterol synthesis, inhibition of its secretion nor changes in cholesterol transport to plasma membrane, esterification of plasma membrane cholesterol and cholesteryl ester hydrolysis. Acyl-CoA:cholesterol acyltransferase (ACAT) activity in microsomes from BFA-treated cells was 1.5- to 1.8-fold higher than that from control cells. The effect of BFA was diminished by treatment with low temperature, which is known to abolish BFA effect on Golgi formation.