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. 2002 Dec;13(12):4414–4428. doi: 10.1091/mbc.E02-02-0116

Figure 5.

Figure 5

Characterization of the secretory pathway in the elo3Δ erg6ts mutant. (A) Secretion of invertase was analyzed in the elo3Δ erg6Δ double mutant strain bearing a plasmid-borne copy of either ERG6 (elo3Δ ERG6, YRS1520) or erg6ts (elo3Δ erg6ts, YRS1519). Cells were preshifted to 37°C for 15 min before invertase induction by glucose limitation. Samples were removed at the time points indicated and the activity of internal and secreted invertase was determined. (B) Maturation of CPY was analyzed in the elo3Δ erg6Δ double mutant strain bearing a plasmid-borne copy of either ERG6 (elo3Δ ERG6, YRS1520) or erg6ts (elo3Δ erg6ts, YRS1519). Cells were preshifted to 37°C for 15 min before labeling with [35S]methionine for 5 min and chasing for the time indicated. CPY was recovered by immunoprecipitation, subjected to SDS-PAGE, and visualized by autoradiography. The position of precursor (p1, ER; p2, Golgi) and mature CPY (m, vacuolar) is indicated. (C) Maturation of Gas1p was analyzed in wild-type (FY1679a), elo3Δ (YRS1118), and the elo3Δ erg6Δ double mutant strain bearing a plasmid-borne copy of either ERG6 (elo3Δ ERG6, YRS1520) or erg6ts (elo3Δ erg6ts, YRS1519). Cells were incubated at 24°C or preshifted to 37°C for 15 min before pulse-chase analysis and immunoprecipitation. The positions of ER precursor (p, 105 kDa) and mature Gas1p (m, 125 kDa) are indicated. Quantification of the degree of Gas1p maturation is shown below.