Fatty acid composition of phospholipids and steryl esters from four nystatin-resistant mutants of Saccharomyces cerevisiae was compared to that from the wild strain. All the mutant strains which produce several ergosterol intermediates incorporated two- to three-fold as much medium chain fatty acids, especially 14:1 in phospholipids, and 12:0, 14:0 and 14:1 in steryl esters as the wild strain did. The increase in the relative amount of medium chain fatty acids in these mutants was found at all the growth temperatures and the growth phases examined, and in all the phospholipid species.