FIGURE 6.

Yet-Sec complex interaction is diminished by treatments that purge translocation substrates. A, semi-intact cells were produced from untagged (CBY2615) or Yet3p-HA (CBY2647) strains, which were treated with or without 0.1 mg/ml of cycloheximide (CHX) 10 or 30 min prior to harvesting. HA immunoprecipitations were performed and analyzed as described in the legend to Fig. 3B. Quantification data from immunoblots are plotted as the mean of the ratio of immunoprecipitation over input, normalized to the no CHX condition (relative IP efficiency). B, semi-intact cells from untagged (CBY740) or Yet3p-HA (CBY2647) strains were washed and incubated with or without an ATP regeneration system as described under “Experimental Procedures” prior to HA immunoprecipitation as in A. Immunoblot data are plotted as the mean of the ratio of +ATP IP efficiency over −ATP IP efficiency (as in Fig. 5), normalized to an IP efficiency of unity for Yet3p-HA. Error bars in both A and B represent the standard deviation with n = 3. For A, p values of <0.05 were achieved between Yet1p and Sec63p, and Yet1p and Sec61p at the 30-min time point (Student's t test, two tailed, paired).