Research Paper Volume 8, Issue 4 pp 810—830

Figure 6. Mutation of the highly conserved K373 in Fkh1 stabilizes the protein in mitosis. (A) Sequence alignment of Fkh1-like proteins across evolutionary boundaries highlighting the highly conserved lysine (K373). The mutation made is shown above in red. (B) Morphology of fkh2Δ cells harboring the FKH1^K-R mutation shown in (A). (C) fkh2Δ cells expressing FKH1^K-R were arrested in mitosis using nocodazole, then treated with CHX for 60 minutes. Proteins were then harvested and examined for Fkh1-TAP, Clb2 and GAPDH content. (D) All bands in (C) were imaged from three separate experiments and the ratio of intensities before and after CHX addition were determined. Standard error of the mean is shown. * - p>0.001. (E) fkh2Δ cells expressing FKH1^K-R were arrested in S phase using HU, then released back into the cell cycle. Samples were removed every 30 minutes afterwards, treated +/− CHX for 1 hr, then proteins were recovered for Westerns. Bands from two experiments were scanned and the ratio of bands +/− CHX was plotted. See also Figure S3.