Figure 3.
HSP104 RNA surveillance characteristics of TRAMP and nuclear exosome components. (A) HSP104 RNA-FISH on fixed samples of the indicated yeast strains after a temperature shift to 37°C for 15 min. HSP104 RNA was detected as described in legend to Figure 2A. DNA was stained with DAPI. The fraction of cells with detectable HSP104 RNA dot signals are shown to the right of the images. For experiments involving the pTET-repressible MTR4 construct, cultures were either treated with 10 μg/ml of doxycycline, or not, for 6 h before the temperature shift. (B) HSP104 Northern (left) or quantitative RT–real-time PCR analysis (right) of RNA harvested from the indicated yeast strains after a 37°C temperature shift for 15 min. Experimental details are as described in legends to Figure 1B and C. The strains mft1Δ/rrp47Δ#1 and mft1Δ/rrp47Δ#2 as well as mft1Δ/air1Δ/air2Δ♯1 and mft1Δ/air1Δ/air2Δ#2 represent two independent spores arising from the relevant crosses.