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

FIGURE 7: Hcm1 abundance is cell cycle dependent and cells are arrested at late M phase upon Ubc1 deletion.

(A) α-factor arrest release of the WT yeast strain harboring an endogenous Hcm1-GFP tag, with detection of Hcm1-GFP and endogenous Clb2. Equal cell numbers were taken after release from G1 phase at the time points indicated, and Western analysis of the cell lysates as shown.

(B) Corresponding FACS analysis of WT samples in (A) at the indicated time points, with unreplicated (1n) and replicated (2n) DNA content indicated. Light images (100x objective) indicate the cell morphology and nuclear position (propidium stained nucleic acid).

(C) RT-PCR (26 cycles) of SNF1 and rRNA expression in the ubc1Δ apc5CA strains, with comparison to isogenic WT.

(D) Flow cytometry of early logarithmic asynchronous cultures demonstrating the relative population of cells with replicated DNA (2n) in ubc1Δ and apc5CA strains, compared to isogenic WT, with representative images of cell morphology and nuclear position. Asy: asynchronous.

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