The structural determinants of checkpoint activation
The structural determinants of checkpoint activation
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DOI：
10.1101/gad.1522607
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发表时间：
2007-04-15
影响因子：
10.5
通讯作者：
Cimprich, Karlene A.
中科院分区：
文献类型：
--
作者：
MacDougall, Christina A.;Byun, Tony S.;Cimprich, Karlene A.
在这里，我们证明了启动的，单链DNA(单链DNA)是足够的激活依赖ATR的检查点途径在非洲爪哇卵提取液。利用这种结构，我们定义了当复制被阻止和正在进行时，5‘和3’端对Chk1激活的贡献。此外，我们还发现，虽然单链DNA不足以激活检查点，但与引物相邻的单链DNA的数量会影响Chk1的磷酸化水平。这些观察定义了检查点激活的最低DNA要求，并表明启动的单链DNA代表了在许多类型的损伤后形成的共同的检查点激活结构。
Here, we demonstrate that primed, single-stranded DNA (ssDNA) is sufficient for activation of the ATR-dependent checkpoint pathway in Xenopus egg extracts. Using this structure, we define the contribution of the 5'- and 3'-primer ends to Chk1 activation when replication is blocked and ongoing. In addition, we show that although ssDNA is not sufficient for checkpoint activation, the amount of ssDNA adjacent to the primer influences the level of Chk1 phosphorylation. These observations define the minimal DNA requirements for checkpoint activation and suggest that primed ssDNA represents a common checkpoint activating-structure formed following many types of damage.