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The toxicity of inorganic metal species towards Saccharomyces cerevisiaehas been shown to be markedly dependent on cellular fatty acid composition. In this investigation, the influence of fatty acid supplementation on the toxicity of the lipophilic organometal, tributyltin was investigated. Growth of S. cerevisiaewas increasingly inhibited when the tributyltin concentration was increased from 0 to 10 μM. However, the inhibitory effect was partly alleviated by supplementation of the medium with 1 mM linoleate (18:2), a treatment that leads to large‐scale incorporation of this polyunsaturated fatty acid (to >60% of total fatty acids) in yeast membrane lipids. Cells that were previously enriched with 18:2 also showed reduced loss of vitality compared to cells grown in the absence of a fatty acid supplement, when exposed to tributyltin. For example, addition of tributyltin to a concentration of 0.1 μM was associated with an approximate 10% reduction in the H+efflux activity of 18:2‐enriched cells, but a 70% reduction in that of fatty acid‐unsupplemented cells. Despite the increased tributyltin resistance of 18:2‐enriched S. cerevisiae,the level of cell‐associated tributyltin was found to be approximately two‐fold higher in these organisms than in fatty acid‐unsupplemented cells. These results demonstrate an increased resistance of 18:2‐enriched membranes to the direct toxic action(s) of tributyltin. This is in contrast to the previously reported effect of 18:2 enrichment on sensitivity of S. cerevisiaeto inorganic metal cations.