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. Author manuscript; available in PMC: 2008 Oct 12.
Published in final edited form as: Mol Cell. 2007 Oct 12;28(1):134–146. doi: 10.1016/j.molcel.2007.09.009

Figure 6. Cell Cycle Control of Ctp1 Abundance.

Figure 6

(A) Transcription of ctp1+ is regulated by MBF. Wild type and nrm1Δ strains were synchronized in G2 by centrifugal elutriation. Samples were taken as cells underwent mitosis and septation. The septation index approximately coincides with S phase. Transcript levels from ctp1+ and cdc22+ were determined by real-time PCR, normalized to act1+ transcript levels and shown as relative transcript levels (%) to maximum wild type levels during the cell cycle (wild type maximum is 100%).

(B) MBF localizes at the ctp1+ promoter region. Association of Cdc10-HA, Res2-HA and Nrm1-HA with the ctp1+, cdc22+ and act1+ promoters was determined by ChIP using log phase cultures. Tagged constructs were expressed from the endogenous loci. Whole cell extract (WCE) from the “no tag” strain was used as a control. Immunoblots confirmed that Nrm1 protein is expressed in res2Δ cells (data not shown).

(C) Immunoblot of Ctp1-TAP in asynchronous, irradiated and HU-treated cells. Immunoblot of Cdc2 with PSTAIR antisera is the loading control.

(D) Immunoblot of Ctp1-TAP in asynchronous, nitrogen-starved or carbon-starved cells. Immunoblot of tubulin is the loading control.

(E) Immunoblot of Ctp1-TAP in cells released from an HU arrest. Septation index indicates completion of the cell cycle.