Fig. 7.

Time-course experiments to analyze re-distribution of p33 from the cytosol to the peroxisomes after shifting down the temperature from nonpermissive to permissive in ssa1^ts yeast. (A) Confocal laser microscopy shows the localization of YFP-p33 in ssa1^ts yeast grown at 23°C in a media containing glucose (to shut down the production of new p33/p92/repRNA) until the shown time point after switching from 32°C. See the details about yeast culturing in the legend to Fig. 6A. Note that the sizes of the characteristic punctate structures containing YFP-p33 grew over time in ssa1^ts yeast cells. (B) Localization of YFP-p33 in SSA1^wt yeast grown at 23°C in a media containing glucose until the shown time point after switching from 32°C. See further details in panel A. (C) Localization of YFP-p33 in SSA1^wt yeast grown continuously at 23°C in a media containing galactose. (D) Western blot analysis of fractionation experiments shows the increasing extent of re-localization of p33 and p92^pol from the soluble fraction to the membrane fraction in ssa1^ts yeast after shifting down the temperature to 23°C. See the details for yeast culturing in panel A. The fractions were obtained as described in Fig. 6B. (E) Western blot analysis of p33 level in total protein extracts prepared from yeast grown as described in panel A and D.