An origin-deficient yeast artificial chromosome triggers a cell cycle checkpoint

被引:44
作者
van Brabant, AJ [1 ]
Buchanan, CD [1 ]
Charboneau, E [1 ]
Fangman, WL [1 ]
Brewer, BJ [1 ]
机构
[1] Univ Washington, Dept Genet, Seattle, WA 98195 USA
关键词
D O I
10.1016/S1097-2765(01)00216-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Checkpoint controls coordinate entry into mitosis with the completion of DNA replication. Depletion of nucleotide precursors by treatment with the drug hydroxyurea triggers such a checkpoint response. However, it is not clear whether the signal for this hydroxyurea-induced checkpoint pathway is the presence of unreplicated DNA, or rather the persistence of single-stranded or damaged DNA. In a yeast artificial chromosome (YAC) we have engineered an similar to 170 kb region lacking efficient replication origins that allows us to explore the specific effects of unreplicated DNA on cell cycle progression. Replication of this YAC extends the length of S phase and causes cells to engage an S/M checkpoint. In the absence of Rad9 the YAC becomes unstable, undergoing deletions within the origin-free region.
引用
收藏
页码:705 / 713
页数:9
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