Back to article: Ydj1 governs fungal morphogenesis and stress response, and facilitates mitochondrial protein import via Mas1 and Mas2

FIGURE 6: The MPP Mas1 and Mas2 are required for mitochondrial function. (A) Assessment of the conditional mutants for MAS1 and MAS2. MAS1 and MAS2 are depleted using the tetracycline repressible promoter. C. albicans wild-type (WT), tetO-MAS1/mas1D and tetO-MAS2/mas2D cells were grown in the absence (No Dox) or presence (+ Dox) of 20 µg/ml doxycycline for 24 hours and then subcultured in the same conditions for 6 hours; transcript levels of MAS1 and MAS2 were measured and normalised to the ACT1 loading control. (B) Depletion of MAS1 or MAS2 reduces C. albicans growth. The tetO-MAS1/mas1D and tetO-MAS2/mas2D strains were grown in the absence (No Dox) or presence (+ Dox) of 20 µg/ml doxycycline for 24 hours, before being subcultured under the same conditions. Growth was measured for a further 8 hours and OD600 was plotted against strains in the absence of depletion. (C) Mas1 is required for oxidative stress survival. The impact of oxidative stress on the tetO-MAS1/mas1Δ and tetO-MAS2/mas2Δ strains and the respective wild type was determined by measurement of CFUs after a 1 hour stress with 5 mM H2O2. Percent CFUs was determined rela-tive to untreated cells. Students paired, two-tailed t-test, * p < 0.05. (D) Depletion of either Mas1 or Mas2 leads to res-piratory growth defects. The WT (SN95), tetO-MAS1/mas1 and tetO-MAS2/mas2 strains were grown in the absence (-) or presence (pre-dox) of 20 μg/ml doxycycline in YPD for 24 hours. The cells were then serially diluted (with the lowest dilution of A600=1) and plated on YP-based plates containing various carbon sources with (+ Dox) or without (- Dox) doxycycline and incubated at 30oC or 37oC for 36 hours.

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