The Saccharomyces cerevisiae strains used are all BY4741 derivative and are listed in Table S1 . To generate cdh1∆::HIS3 strain, cdh1∆::KanMX4 was transformed with a DNA fragment containing HIS3MX. To construct double mutant strains, the DNA fragment containing cdh1∆::HIS3 was amplified and transformed in the deletion strains. To generate Rpn4-HAcdh1Δ::kan strain, Rpn4-HA:HIS3 was transformed with a DNA fragment containing cdh1∆::KanMX4. Strains were transformed by the standard lithium acetate procedure [59 (link)]. Gene deletion was confirmed by PCR. For overexpression of Cdh1-m11 and Sod2p, cells were transformed with the plasmids pRS416-GALL-3HA-Cdh1-m11 [26 (link)] and Yep352-SOD2 [45 (link)], respectively.
Cells were grown in rich medium [YPGal: 2% (w/v) galactose, 1% (w/v) yeast extract, 2% (w/v) bactopeptone] or synthetic complete medium [SC: 0.67% (w/v) Bacto-yeast nitrogen base w/o amino acids, 2% (w/v) glucose and 0.2% (w/v) Dropout mix] lacking uracil/leucine, as appropriate. For Cdh1-m11 overexpression, cells were grown in YPRaff medium [2% (w/v) raffinose, 1% (w/v) yeast extract, 2% (w/v) bactopeptone] overnight until mid-log phase and cultured with 4% galactose for 3h before oxygen consumption analysis. Cultures were routinely grown at 26 °C in an orbital shaker at 140 r.p.m.
Cells were grown in rich medium [YPGal: 2% (w/v) galactose, 1% (w/v) yeast extract, 2% (w/v) bactopeptone] or synthetic complete medium [SC: 0.67% (w/v) Bacto-yeast nitrogen base w/o amino acids, 2% (w/v) glucose and 0.2% (w/v) Dropout mix] lacking uracil/leucine, as appropriate. For Cdh1-m11 overexpression, cells were grown in YPRaff medium [2% (w/v) raffinose, 1% (w/v) yeast extract, 2% (w/v) bactopeptone] overnight until mid-log phase and cultured with 4% galactose for 3h before oxygen consumption analysis. Cultures were routinely grown at 26 °C in an orbital shaker at 140 r.p.m.
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