JCS005314 Supplementary Material

Files in this Data Supplement:

• Supplemental Table 1 -

  Adobe PDF

• Supplemental Figure 1 -

  Fig. S1. FACS analysis of cdk5^ts cells upon shift to the restrictive temperature (34°C). Wild-type (FB1) and cdk5^ts cells were grown in liquid CMD medium until OD₆₀₀: 0.02 at 22°C. Half of the culture was incubated at 34°C and the rest maintained at 22°C. Samples were taken for FACS analysis at the indicated times. Peaks below 1C DNA content were the result of cell debris fluorescence after cell lysis. The 2C DNA content peak in cdk5^ts cells growing at 34°C corresponds mainly to cell doublets.

• Supplemental Figure 2 -

  Fig. S2. Reversible phenotype of the cdk5^ts mutant. cdk5^ts mutant cells grown in CMD medium at 22°C were transferred to 34°C. After incubation for 6 hours, the cells were shifted down to 22°C and incubated for 10 hours. The cells at the indicated times were taken for observation of cell morphology after having been stained with Calcofluor. Quantification of the different morphotypes is shown in the upper part.

• Supplemental Figure 3 -

  Fig. S3. (A) Micrographs show the morphology of cdk5^ts cells growing in liquid YPD at permissive (22°C), semipermissive (30°C) and restrictive temperatures (34°C). (B) Temperature-sensitive growth of cdk5^ts cells in solid medium (YPD) is not suppressed by the addition of 1M sorbitol. (C) Addition of 1M sorbitol did not prevent cell lysis. Images of cdk5^ts cells incubated in YPD with 1M sorbitol at 34°C for 12 hours.

• Supplemental Figure 4 -

  Fig. S4. (A) Cdc24 is essential and is required for polarity in U. maydis cells. UMI20 cells carrying a conditional allele, cdc24^nar, in which the cdc24 was placed under the control of the nar1 promoter (Brachmann et al., 2001) were spotted in permissive (minimal medium with nitrate as nitrogen source) and restrictive (CMD) conditions. (B) Morphology of cdc24^nar cells after 6 hours’ incubation at restrictive conditions (liquid CMD, 28°C). Note that the majority of the cells arrest as ellipsoidal doublets. Also observe the diffuse pattern of calcofluor staining.

• Movie 1 -

  Movie 1. GFP-Myo5 dynamics at the tip in cdk5^ts cells incubated at permissive (22°C) and restrictive (34°C) temperature. Images series of 60 frames at 1-second intervals were taken. Elapsed time is given in seconds:milliseconds. Bar, 2 μm

• Movie 2 -

  Movie 2. GFP-Myo5 dynamics at the tip in cdk5^ts cells incubated at restrictive (34°C) temperature. Images series of 60 frames at 1-second intervals were taken.

• Movie 3 -

  Movie 3. GFP-Myo5 dynamics at the tip in cdk5^ts cells overexpressing cdc24 incubated at restrictive (34°C) temperature. Images series of 60 frames at 1-second intervals were taken.