Figure 5
From: Yeast karyopherin Kap95 is required for cell cycle progression at Start
Control of the subcellular localization of Mbp1. (A) Cells from exponentially growing cultures of the wild type (JCY816) and tetO 7 :KAP95 (JCY848) strains expressing the HA-tagged version of Mbp1, were incubated in the presence of 5 μg/mL doxycycline for 6 hours and assayed by indirect immunofluorescence. The HA indirect-fluorescence signals, the DAPI staining of DNA and DIC images are shown. (B) Putative NLSs sequences in the Mbp1 protein are indicated as white boxes. Amino acids substitutions introduced to inactivate the NLSs are shown in grey. Grey boxes represent the DNA binding (DBD), the ankyrin (ANK) and the Swi6 binding (SBD) domains. (C) Exponentially growing cells of the wild type strain (W303-1a) transformed with plasmids pMBP123-184-GFP4, pMBP123-184NLS1i-GFP4, pMBP123-184NLS2i-GFP4 and pMBP123-184NLS1i2i-GFP4 were analyzed by fluorescence microscopy. GFP signal images are shown. (D) Exponentially growing cells of of the wild type (W303-1a) and tetO 7 :KAP95 (JCY635) strains transformed with plasmid pMBP123-184-GFP4 were incubated in the presence of 5 μg/mL doxycycline for 6 hours and analyzed by fluorescence microscopy. GFP signal and DIC images are shown. (E) Exponentially growing cells of the the wild type (W303-1a) and the srp1 strains transformed with plasmid pMBP123-184-GFP4 were incubated at 37° for 2 hours. GFP signal images are shown.