Article Figures & Data
Figures
- Fig. 1.
mRNA levels of SSB genes upon a carbon upshift. At time zero, glucose was added to wild-type cells growing in glycerol-based medium. At left, signals from Northern blotting of several transcripts upon a carbon upshift are shown. The graph at right shows quantification of the signals after normalization to snRNA U2 levels.
- Fig. 2.
Localization of the 5′ ends of the SSB1 andSSB2 genes and analysis of their mRNA levels upon a carbon upshift. (A) Primer extension reactions are shown in the center: lane 1, strain overexpressing the SSB1 gene; lane 2, wild-type cells; lane 3, strain overexpressing the SSB2 gene. Sequencing reactions are shown on the left for the SSB1 gene and on the right for the SSB2 gene. (B) Sequences from theSSB1 and SSB2 genes showing the start site of transcription relative to the initiation ATG codon. Sequences complementary to the primer are located above the arrow labeled as primer. (C) mRNA levels of SSB1 and SSB2 genes after a carbon upshift. Samples of the culture were collected at the indicated times after glucose addition. At left is shown a sequencing gel which separates the primer extension products. Lane G shows results from cells grown on glucose-based medium. The graph shows the quantification of the signal obtained after normalization to snRNA U4.
- Fig. 3.
mRNA levels of various transcripts after amino acid limitation. 3AT was added to cells (strain F113) growing in SD minimal medium at time zero. RNA samples were analyzed by Northern blotting. (A) (Left) Control from cells that had no 3AT (−3AT) treatment. (Right) Signals obtained from cells treated with 3AT. Genes analyzed were SSB, RPL5 (RP), SSA (a yeast cytosolic Hsp70), and HHO1 (histone H1). HIS3 was used as a control to detect the efficiency of the treatment. (B and C) Quantification of the mRNA levels in the presence of 3AT (+3AT) and ofHIS3 mRNA with (+3AT) and without (−3AT) 3AT, as shown in panel A, after normalization to actin transcript levels.
- Fig. 4.
mRNA levels of ribosomal components upon a heat shock in the wild-type (WT) strain and in cells containing the EXA3-1mutation. JH27A (wild-type) and MH297 (EXA3-1) cells growing in YPD medium at 23°C were rapidly shifted to 39°C. Aliquots of the culture were collected at the times indicated after the temperature upshift, and extracted RNA was subjected to Northern blot analysis. (Top) Northern blot images. (Bottom) Quantification after normalization to rRNA (see Materials and Methods).
- Fig. 5.
Levels of MATα1/2 and URA3transcripts upon a heat shock in the wild-type (WT) (JH27A) strain and in cells containing the EXA3-1 mutation (MH297). Experiments were performed as described for Fig. 4. In the case of the short-half-life message URA3, samples were taken at 0 to 18 min after a temperature upshift (see Materials and Methods). (Top) Northern blot images. (bottom) Graphs showing the quantification of signal after normalization to rRNA.
- Fig. 6.
mRNA levels of an SSB1:URA3 fusion in wild-type (WT) and mutant MH297 cells after a heat shock. The experiment was performed as described for URA3 in Materials and Methods. Northern blot images of each transcript are shown at left. (A) SSB1:URA3 fusion; (B) RPL11. Quantification of the signals detected in panels A and B is shown on the right. Signals were normalized to rRNA levels.
- Fig. 7.
Influence of the EXA3-1 mutation on the transcript levels of a CUP1:SSB1 fusion. The experiment was done as described in the legend to Fig. 5. At left, Northern blot images for CUP1:SSB1 fusion (A) andRPL11 (B) are shown. At right is shown quantification of the signals after a heat shock with rRNA as a loading control.
