Back to article: The ubiquitin-conjugating enzyme, Ubc1, indirectly regulates SNF1 kinase activity via Forkhead-dependent transcription

FIGURE 6. Hcm1 is impaired in its cell cycle dependent nuclear import upon deletion of Ubc1.

(A) Equal numbers of early logarithmic phase cells from WT and ubc1Δ strains with or without an endogenous Hcm1-TAP tag were lysed followed by TAP Western analysis.

(B) Assessment of Hcm1 protein stability over 3 hours in WT and ubc1Δ strains in the presence of cycloheximide, added at time (0), in 2% glucose. Equal cell numbers were removed at the indicated time points with 30 μg protein loaded.

(C) Fluorescent microscopy of genomically integrated GFP-tagged Hcm1 expressed from its endogenous promoter in isogenic WT and ubc1Δ strains. Both strains were arrested in G1 followed by release, with cells collected at the indicated timepoints.

(D) Flow cytometry analysis of cells collected at the timepoints in (A), highlighting the relative proportion of replicated DNA (2n) upon release from G1.

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