Overexpression of ΔC-HookA by the gpdA promoter drives dynein–dynactin relocation from the MT plus ends to the minus ends. (A) A diagram showing the wild-type hookA allele, the ΔC-hookA-S allele, and the gpdA-ΔC-hookA-S allele. (B) Western blots showing the ΔC-HookA-S protein being overexpressed by the gpdA promoter. GFP-histone H1 was used as a loading control. (C) Colony phenotype of a gpdA-ΔC-hookA-S strain in comparison to that of a wild-type control and a ΔC-hookA-S strain. (D) Images of GFP-dynein in wild-type and gpdA-ΔC-hookA-S strains. Plus-end dynein comets near the hyphal tip (arrowheads) are present in wild-type, but not in a gpdA-ΔC-hookA-S hypha, although the plus-end comets of mCherry-ClipA/CLIP-170 are present in the same gpdA-ΔC-hookA-S hypha. Brightness is increased by 30% for all three images. (E) Accumulation of GFP-dynein as plus-end comets in a wild-type control and on a septum (arrows) in a gpdA-ΔC-hookA-S strain. Bright-field images are shown below to indicate hyphal shape and position of the septum. (F) Accumulation of p150-dynactin-GFP as plus-end comets in a wild-type control and on a septum in a gpdA-ΔC-hookA-S strain. (G) The localization of GFP-dynein on nuclei labeled with NLS-DsRed in the gpdA-ΔC-hookA-S strain. A wild-type control is presented. Brightness is increased by 50% for both images. (H) Plus-end dynein comets in the nudAF208V, gpdA-ΔC-hookA-S strain. Bars, 5 µm.
