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. 2002 Jul;22(13):4477–4490. doi: 10.1128/MCB.22.13.4477-4490.2002

FIG. 2.

FIG. 2.

Motif C is not required for the essential function of dfp1+. (A) Schematic of the Dfp1 C-terminal mutants and summary of the dfp1Δ complementation analysis. (B) Plasmid shuffle assay showing complementation of dfp1Δ by dfp1 C-terminal mutants. Transformants were streaked onto YE or YE plus 10 mg of FUdR/liter and grown at 30°C. Colony formation on FUdR indicates complementation. (C) Relative growth of logarithmic-phase cultures of dfp1+ (GBY572), dfp11-459 (AFY6), and dfp11-376 (AFY7) cells was assessed by measuring the optical density of the cultures at the indicated time points. (D) The DNA contents of logarithmically growing cultures of dfp1+ (GBY572), dfp11-459 (AFY6), and dfp11-376 (AFY7) cells were measured by flow cytometry. The control sample (dfp1 depletion, AFY4) was obtained following the repression of dfp1 expression by growth in the presence of 5 μg of thiamine/ml for 4 h. The positions of 1C and 2C DNA contents are indicated. (E) Progression through S phase was examined by flow cytometry. dfp1+ (GBY572) and the motif C mutant dfp11-376 (AFY7) were grown to mid-logarithmic phase (log), blocked in 25 mM HU for 4 h (+HU), and then released into the cell cycle. Cultures were sampled at the indicated times following release from HU. The positions of 1C and 2C DNA contents are indicated.