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. 2009 Feb 6;5(2):e1000294. doi: 10.1371/journal.ppat.1000294

Figure 2. Tor1 regulates the expression of the hyphae-specific transcripts ALS1, ALS3, HWP1, and ECE1 and cellular aggregation via the GPI-anchored proteins Als1 and Als3.

Figure 2

(A) Northern analysis of ALS1, ALS3, HWP1, and ECE1 in wild type and TOR1-1/TOR1 cells grown in Spider media at 37°C and treated with 20 nM rapamycin and vehicle control for 90 minutes. (B) Expression of ECE1 is upregulated in a C. albicans FKBP12 homozygous null mutant strain (rbp1/rbp1) grown under the same conditions as in (A). (C) Rapamycin inhibition of Tor1 results in increased Als3 at the cell wall. Als3 localization was assayed by indirect immunofluorescence using purified anti-Als3p serum in wild type and als3/als3 strains grown in the same conditions as in (A). (I) Wild type cells+vehicle, (II) wild type cells+rapamycin, (III) magnified view of a hyphal cell from (II), (IV) magnified view of yeast and hyphal cells from (II), (V) als3/als3 mutant cells+rapamycin. Yeast cells are indicated by arrows. Red fluor corresponds to Alexa 594 and blue to DAPI. (D) Cells of the als1/als1 als3/als3 double mutant strain grown in Spider media at 37°C fail to aggregate upon treatment with 20 nM rapamycin. Cellular aggregation is restored when this strain is complemented by ectopic expression of either ALS1 or ALS3.