FIG 6.

Physiological significance of the phosphorylation states of Pkc1 at Thr¹¹²⁵ and Ser¹¹⁴³ on the growth of yeast cells. (A) pkc1Δ (DL376) cells carrying various PKC1 alleles (WT, PKC1^WT; T1125A, PKC1^T1125A; S1143A, PKC1^S1143A; T1125A/S1143A, PKC1^{T1125A/S1143A}) with YCp50 (a CEN-type plasmid) were cultured in SD medium until the log phase of growth. Cells were then serially diluted (1:10) with a 0.85% NaCl solution, and 4 μl of each cell suspension was spotted onto SD agar plates containing MG. Cells were grown at 28°C for 3 days. (B) pkc1Δ cells (DL376) carrying various PKC1 alleles (WT, PKC1^WT; ΔHR1, PKC1^ΔHR1; 4C/S, PKC1^4C/S) with pFL39 (a CEN-type plasmid) were streaked onto SD agar plates with or without 1 M sorbitol and cultured at 28°C for 3 days. (C) Cells with GAL1 promoter-driven AVO1 (RL25-1C) carrying various PKC1 alleles (WT, PKC1^WT; T1125A, PKC1^T1125A; S1143A, PKC1^S1143A; T1125A/S1143A, PKC1^{T1125A/S1143A}) with YEp352 (a 2μ-type plasmid) were cultured in SC-Gal medium until the log phase of growth. Cells were then serially diluted (1:10) with 0.85% NaCl solution, and 4 μl of each cell suspension was spotted onto SC-Gal (Gal) or SD (Glc) agar plates. Cells were cultured at 28°C for 3 days.