Fig. 1.
Cells’ requirement of VLCFA is significant at elevated temperature. (A) BY4741 (wild-type), 5281 (sur4Δ), 5763 (fen1Δ), YOK2230 (PtetO7-PHS1), MRY9 (PtetO7-PHS1 sur4Δ), and MRY7 (PtetO7-PHS1 fen1Δ) cells were grown to stationary phase, serially diluted at 1:10, spotted on YPD plates, grown for the indicated times at 30°C or 39°C, and photographed against a dark background. (B) YOK2230 (PtetO7-PHS1) and MRY9 (PtetO7-PHS1 sur4Δ) cells were grown to log phase at 30°C, and total membrane fractions were prepared. Samples (20 μg) were incubated with 20 μM palmitoyl-CoA, 73 μM malonyl-CoA, and 27.3 μM [14C]malonyl-CoA (0.075 μCi) for 1 h at 37°C. Lipids were saponified, acidified, extracted, converted to methyl ester forms, separated by reverse-phase TLC, and detected by autoradiography.