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Figure 8


Fig. 8. Bem1 has a functional NES. (A) Bem1 protein has a putative nuclear export sequence (NES). Point mutations to destroy this NES were introduced in Bem1 creating Bem1-NES{Delta}. The NES of PKI was cloned in Bem1-NES{Delta} resulting in Bem1-NES{Delta}+NES. These three constructs were tagged at the C-terminus with GFP. (B) bem1-GFP (UMI64), bem1-NES{Delta}-GFP (UMI136) and bem1-NES{Delta}+NES-GFP (UMI137) cells were grown in CMD. Bem1-GFP localizes at the cell tips. Bem1-NES{Delta}-GFP accumulates within the nucleus and is also observed in the cytoplasm. A faint signal of Bem1-NES{Delta}+NES-GFP is distributed in the cytoplasm without any specific accumulation. Note that bem1-NES{Delta}-GFP cells presented loss of polarity but this was rescued by introducing the extra PKI NES (bem1-NES{Delta}+NES-GFP cells). Scale bar: 10 µm.