Figure 6.
Intracellular targeting by PH‐like domain of Vps13p. A‐F, Vps13‐EGFP constructs, A/B: wildtype (WT), C/D: L3125A I3129A (LIAA) and E/F: 1–3028 (ΔPH) in Δvps13 cells in log phase (A/C/E) or early stationary phase (B/D/F). Sites of localization include intracellular puncta (filled arrowheads) in log phase, and the nucleus vacuole junction (NVJ, open arrowheads) in stationary phase. Scale bar (shown in A only) 5 µm. G, Overlay blots to detect secreted CPY. Controls: wild‐type cells and Δvps13 cells show no secretion and maximal secretion, respectively. Rescue of Δvps13 was tested for Vps13‐EGFP plasmids as in A‐F. Results are representative of 6 similar experiments. H, Residue conservation in the C‐terminal 1095 residues of Vps13p, calculated by ConSurf and scaled as described in Materials and Methods. The most conserved part of the Vps13 C‐terminus is a glycine rich domain (orange) of no known function,72 not the predicted PH domain (green).