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. 2007 Oct;18(10):4155–4167. doi: 10.1091/mbc.E07-02-0094

Figure 7.

Figure 7.

Bud6p targets for3p to cell tips through both anchoring and relief of autoinhibition. (A) Mapping of the BBS in for3p by two-hybrid analysis. Interaction of C-terminal for3p fragments cloned in the pGBD vector with pGAD-bud6C(581-1385) was assayed by growth on −His plates. Note how the minimum interaction domain overlaps with the DAD region. (B) Single focal plane widefield fluorescent images of for3p-3GFP in bud6Δ cells. Cells on the right were treated with 200 μM LatA. (C) Single focal plane widefield fluorescent images of for3pDAD*-2GFP bud6Δ cells. Cells on the right were treated with 200 μM LatA. Note that the DAD* mutation is sufficient to restore efficient for3p localization to cell tips in bud6Δ mutant cells (blue arrowheads). (D) Projection images of spinning disk confocal stacks of GFP-for3N (left) and GFP-for3C (middle), and GFP-for3C-I930A (right) expressed from the medial-strength nmt1 promoter in bud6Δ mutant cells. GFP-for3C fails to localize to cell tips. (E) Single focal plane widefield fluorescent images of cdc42p-GFP in wild-type and bud6Δ cells. (F) Single focal plane widefield fluorescent images of bud6p-GFP in wild-type and cdc42-1625 cells.