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. 2010 Oct 29;285(53):41244–41254. doi: 10.1074/jbc.M110.155242

FIGURE 3.

FIGURE 3.

Dbf4-RSIEGA mutants suppress the cdc5-1 temperature sensitivity. A, the indicated strains W303-1A, dbf4-Δ82–88 (M2805), cdc5-1 (M1614), cdc5-1 dbf4-Δ82–88 (M3112 and M3114), cdc5-1 dbf4-R83E (M3116, M3117), and cdc5-1 dbf4-NΔ109 (M2655, M2656) were spotted onto YPD plates and scored for growth at the indicated temperatures. B, various dbf4 deletions on an ARS CEN plasmid (pMW489) were introduced into M2600 (cdc5-1 dbf4Δ::kanMX6) and scored for growth by spotting serial dilutions on YPD media at the indicated temperatures. C, shown is a summary of dbf4 mutations, their effect on the Dbf4-PBD interaction, and suppression of the cdc5-1 ts. dbf4 mutants were scored for growth in the M2600 (cdc5-1 dbf4Δ::kanMX6) background by spotting serial dilutions on YPD media at increasing temperatures (supplemental Fig. S4). D, high copy plasmids expressing wild type DBF4, and the indicated mutants were transformed into M1614 (cdc5-1). Cultures were spotted onto SCM−Leu plates at 25 °C, indicating that high copy dbf4-NΔ65 lethality is alleviated by deleting residues 82–88. E, expression of the Dbf4 N terminus from the GAL1,10 promoter is lethal to cdc5-1 cells only if Dbf4 retains the ability to interact with Cdc5 as occurs in the WT, S84A, S84E, E86A, and E86K mutants.