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Fig. 7. yvh1 is a suppressor of pma1-10, causing cell-surface stability of Pma1-10. (A) Cells were grown at 30 and 37°C on plates with selective synthetic complete medium. (B) Pulse-chase analysis of Pma1-10 in YVH1 (XGY32) and yvh1 ${Delta}$ (KKY11) cells. Cells were pulse-labeled with Expre³⁵S³⁵S for 5 minutes and chased for various times. Immunoprecipitations with anti-Pma1 antibody were normalized to acid-precipitable cpm. (C) Endocytosis in yvh1 cells. Cells bearing pGAL-myc-STE3 (pSL2015) were grown in galactose-containing medium to induce expression of myc-Ste3. At various times after shifting to glucose, cells were lysed and myc-Ste3 was analyzed by western blot. (D) Pma1-10 is delivered to the plasma membrane in yvh1 cells. pma1-10 yvh1 (KKY11) and pma1-10 pep4 (XGX28-1A) cells were pulse-labeled for 5 minutes and chased for 1 hour at 37°C. Lysate was fractionated on Renografin density gradients. Newly synthesized Pma1-10 was immunoprecipitated from pooled fractions and analyzed by SDS-PAGE and autoradiography. Distribution of the cell-surface marker Gas1 was analyzed by western blotting of pelleted membranes. (E) Electrophoretic mobility of newly synthesized Pma1. Wild-type, yvh1 pma1-10, and pep4 pma1-10 cells were pulse-labeled and chased for 2 hours at 37°C. Pma1 was immunoprecipitated with anti-Pma1 antibody and analyzed by SDS-PAGE and autoradiography.

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