Fig. 8. Mutation of a C-terminal KY motif affects bud neck localization of Clb2. (A) A GFP tag was appended to the clb2^K459A,Y460C allele at the CLB2 locus. Cells were processed for microscopic observations as described in Fig. 1 and are shown in comparison with CLB2-GFP cells before (a) and during (b-d) nuclear division. (B) The indicated mitotic cyclins were overexpressed as GFP fusion under the control of the GAL1 promoter. Cells grown in SC-raffinose were collected on nitrocellulose filters and induced for 2 hours on YEP-galactose plates before microscopic observation. (C) The percentage of cells with Bem1-GFP polarized at the bud tip was monitored in clb2^K459A,Y460C and clb2^{K459A,Y460C,NLS} cells as described in Fig. 5. Bar, 2µm.