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Fig. 6. Requirements for Cdr2p localization. (A) Schematic diagram of cdr2 deletion constructs. The black box indicates the N-terminal kinase domain; the asterisk indicates the D133A mutation. Cells expressing each construct were tested for their localization pattern and their ability to complement the nitrogen deprivation defect of cdr2 null cells; see Fig. 7. (B-H) Wild-type cells (KGY246) expressing (B) GFP-Cdr2p or (C) GFP-Cdr2p(E177A); cdr2 cells (KGY520) expressing (D) GFP-Cdr2p(747-775) or (E) GFP-Cdr2p(1-252) under the control of the weak nmt81 promoter; cdr2 cells (KGY520) expressing (F) Cdr2pE177A, (G) Cdr2p(1-252), or (H) Cdr2p(332-775) under the control of the strong nmt1 promoter. All cells were grown in the absence of thiamine for 18 hours at 32°C. Images of live cells were captured in B-E. Cells shown in F-H were fixed in 10% formaldehyde solution and stained with calcofluor.





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