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. 2013 Jul 18;12(16):2564–2569. doi: 10.4161/cc.25692

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Copyright © 2013 Landes Bioscience

This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited.

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graphic file with name cc-12-2564-g2.jpg — Figure 2. Regulation of Mrc1 cellular amounts during replication and checkpoint. (A) mrc1-S860A/S864A mutation stabilizes Mrc1. Exponentially growing cells of the indicated genotypes were treated with 0.1 mg/ml cycloheximide at 30 ˚C. Cellular amounts of Mrc1-FLAG and Mrc1-S860A/S864A-FLAG were examined at various time points between 0 and 4 h of cycloheximide treatment. The anti-FLAG (M2) antibody was used to detect Mrc1. Western blotting of tubulin was performed as a loading control. (B) Models of phosphorylation-dependent Mrc1 regulation during replication and checkpoint. Pof3 controls replisome quality via ubiquitination/degradation of Pol2 when the fork is adversely blocked and the checkpoint is activated, while Pof3 also regulates Mrc1 to deactivate checkpoint when the replisome is intact to resume and complete DNA replication.