Fig 1.
TAT2/Tryptophan rescues SDS sensitivity in Δmck1 cells.
(A) W303 MCK1 cells (his3-11,15 leu2-3,112 trp1-1 ura3-1) and isogenic Δmck1 cells transformed with TAT2/pRS425 plasmid were 5-fold serially diluted onto YPD or YPD plus 0.0075% SDS. (B) The same strains, MCK1 or Δmck1, used in A were struck on YPD or YPD containing 0.0075% SDS and supplemented on top with excess tryptophan (+TRP), adenine (+ADE), leucine (+LEU), uracil (+URA) or histidine (+HIS). (C) The indicated strains were 10-fold serially diluted onto YPD or YPD plus 0.0075% SDS and (D) containing additional tryptophan (+TRP), tyrosine (+TYR), phenylalanine (+PHE) or histidine (+HIS).
Fig 2.
TRP1 prototrophy confers growth advantage in the presence of SDS.
(A) The indicated cells were 10-fold serially diluted onto YPD or SD plates with or without 0.0075% SDS. (B) Log phase cells of the genotype ade2-1 his3-11,15 leu2-3,112 trp1-1 ura3-1 (trp1-1) and isogenic TRP1 cells (TRP1) were grown in YPD with or without SDS (0.001%). Every hour for 4 hours the optical density (OD) was read and an aliquot of cells was stained with propidium iodide and analyzed by FACS. The bottom row is an example of the FACS readings at the 4 hour time-point and the gating used to determine live vs dead cells. The graph on the left indicates the percentage of dead cells. The graph on the right indicates the doubling times as the slope of the line determined by plotting log2 of the optical density reading vs. time.
Fig 3.
TRP1 prototrophy recovers growth sensitivity due to some cell wall/membrane damaging treatments but not all.
The indicated yeast cells were 10-fold serially diluted onto YPD (A) with or without 0.0075% SDS, (B) containing 10ug/ml Calcofluor White or 10ug/ml Congo Red, (C) incubated at 37°C or 39°C or (D) containing 0.15% Tea Tree Oil.
Fig 4.
Cells deficient in the biosynthesis of tryptophan or tyrosine are sensitive to SDS.
(A) Schematic of the tryptophan, phenylalanine and tyrosine biosynthesis pathway. (B and C) W303 cells with TRP1 (ade2-1, his3-11,15, leu2-3,112 and ura3-1) are labeled as ahlTu. W303 cells harboring specified deletions in the tryptophan, phenylalanine and tyrosine biosynthesis pathway were 10-fold serially diluted onto YPD or SD with or without SDS.
Fig 5.
SDS enhances tryptophan uptake.
The import rates of radiolabeled tryptophan and histidine were measured using W303 prototrophic cells (ADE2, HIS3, LEU2, TRP1, and URA3) in the absence (squares) or presence of (open circles) 0.0075% SDS or (solid circles) 0.005% SDS (See Materials and methods). Shown are representative curves.