Figure 8.

The PER17/BST1 and ERV29 genes are required for the degradation and transport of overexpressed CPY*. (A) and (B) Wild-type (ESY342) and per17/bst1 (ESY347) cells overexpressing CPY* were subjected to pulse-chase analysis. CPY* was immunoprecipitated from detergent lysates and divided into two aliquots. From one CPY* was released from protein A resin and immunoprecipitated using anti-α 1,6-mannose antibodies. Immunoprecipitated proteins were treated with Endo H, and resolved by SDS-PAGE. Quantitative analysis was performed using a phosphorimager and plotted in B. The fraction of CPY* gaining α 1,6-mannose addition was determined by quantifying the ratio of CPY* immunoprecipitated with anti-α 1,6 antibody vs. the CPY* immunoprecipitated using anti-HA antibody. (C) and (D) Wild-type (ESY342) and Δerv29 (ESY386) cells overexpressing CPY* were subjected to pulse-chase analysis as in A and B.