Figure 7.

Functional and biochemical links between Arl1p and Gga2p. (A) The subcellular localization of HA–Gga2p depends on Arl1p. Indirect immunofluorescence was performed with untagged (control), wild-type (wt), Δarl1, vps27, and Δarl1 vps27 cells expressing HA–Gga2p; ^*the expanded class E compartment. Bar, 5 μm. (B, C) Interactions between GST–Gga2 fusions and Arlp and Gga2p in vitro. Yeast cell lysates with HA–Arl1p and Myc–Gga2p were preincubated for 10 min at 30°C with either 0.5 mM GTP, GDP, or GTP-γ-S and incubated with the indicated matrix-bound GST fusions. Bound proteins were analysed as described in Figure 4B using anti-HA and anti-c-Myc antibodies.