Figure 5.—

Overproduced WT and A53T are toxic in yeast, while A30P is not. (A) Y382 or TW373 cells carrying the indicated GST fusions were grown to stationary phase. Tenfold serial dilutions were spotted onto synthetic medium lacking uracil and containing 2% glucose (dextrose) (SD −Ura) or synthetic medium lacking leucine and containing 2% galactose (SGal −Leu) and incubated at 30° for 3 days. (B) Y382 cultures expressing the indicated GST fusions were grown to midlog phase in −Leu medium and shifted to galactose for 3 hr to induce expression of the indicated fusion proteins. Lysates were prepared and subjected to immunoblot analysis using an anti-GST antibody. The blot was then reprobed with an anti-Cdc34 antibody as a loading control.