Figure 7. Pom1 redistribution under glucose restriction disrupts Wee1 regulation at cortical nodes.

(A) Localization of Wee1-mNG in wild-type or pom1∆ cells grown in either high (2%) or low (0.03%) glucose media. Images were collected using TIRF microscopy. Scale bar, 5 µm. (B) Quantification of Wee1 bursting kinetics in wild-type cells grown under high (2%, blue circles) and low (0.03%, red squares) glucose conditions. The top panel is a plot of the total number of Wee1 bursts as a function of cell length, counted in single time point TIRF micrographs. The middle panel is a plot of the frequency of Wee1 bursts as a function of cell length. The bottom panel is a plot of Wee1 burst duration as a function of cell length. (C) Quantification of Wee1 bursting kinetics in pom1∆ cells grown under normal (2%, green circles) and low (0.03%, magenta squares) glucose conditions, as in panel B. For (B–C), insets represent whether the best fit values of the slope (b₁) and/or Y-intercept (b₀) of the two linear regressions are different at the 95% confidence level. See Supplementary file 1 and Supplementary file 2 for parameters of linear regressions and full statistical comparisons. (D) Western blot of Wee1 in wild-type, pom1∆, and cdr2∆ cells grown in high (2%) or low (0.03%) glucose media showing phosphorylation dependent band-shifts.

Figure 7—figure supplement 1. Wee1 bursting in low glucose phenocopies a kinase-dead Cdr2 mutant.

(A) Plot of the frequency of Wee1 bursts as a function of cell length in wild-type cells grown in high glucose (blue circles) and low glucose (red squares), or kinase-dead cdr2(E177A) mutant cells grown in high glucose (green triangles). (B) Plot of Wee1 burst duration as a function of cell length in wild-type cells grown in low glucose (red squares) and kinase-dead cdr2(E177A) mutant cells grown in high glucose (green triangles). For (A–B), insets represent whether the best fit values of the slope (b₁) and/or Y-intercept (b₀) of the linear regressions are different at the 95% confidence level. See Supplementary file 1 and Supplementary file 2 for parameters of linear regressions and full statistical comparisons.