Alteration of ROS Homeostasis and Decreased Lifespan in S. cerevisiae Elicited by Deletion of the Mitochondrial Translocator FLX1
Figure 1
(a) Respiratory-deficient phenotype of flx1Δ strain: effect of succinate and malate addition. WT and flX1Δ cells were cultured at 30°C in YEP liquid medium supplemented with either glucose or glycerol (2% each) as carbon source. Where indicated either 5 mM succinate (Succ) or 5 mM malate (Mal) was added. Cell growth was estimated at the stationary phase (24 h) by measuring the absorbance at 600 nm () of a ten-fold dilution of each growth culture, consistently, corrected for the dilution factor. The values reported in the histogram are the means (±SD) of three experiments. (b) Changes in the recombinant Sdh1-HAp level in flx1Δ strain. Cellular lysates were prepared from WT-HA and flX1Δ-HA cells grown at 30°C up to the exponential growth phase (5 h) in YEP liquid medium supplemented with either glycerol or galactose (2% each) as carbon source. Proteins from cellular lysates (0.05 mg) were separated by SDS/PAGE and transferred onto a PVDF membrane. In each extract, Sdh1-HA protein was detected by using an α-HA and its amount was densitometrically evaluated. The values reported in the histogram are the means (±SD) of three experiments performed with different cellular lysates preparations. Statistical evaluation was carried out according to Student’s -test (). As a control, the specific activity of the enzyme fumarase (FUM) was determined in each cellular lysate preparation.
