An essential role for Cdk1 in S phase control is revealed via chemical genetics in vertebrate cells.
An essential role for Cdk1 in S phase control is revealed via chemical genetics in vertebrate cells.
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DOI：
10.1083/jcb.200702034
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发表时间：
2007-07-16
影响因子：
7.8
通讯作者：
Takeda, Shunichi
中科院分区：
文献类型：
--
作者：
Hochegger, Helfrid;Deisuphong, Donniphat;Sonoda, Eiichiro;Sciberi, Alihossein;Rajendra, Eeson;Kirk, Jane;Hunt, Tim;Takeda, Shunichi
在脊椎动物中，需要Cdk 1启动有丝分裂;然而，这种激酶在S期的任何功能仍不清楚。为了研究这一点，我们产生了鸡DT 40突变体，其中类似物敏感的突变体cdk 1取代了内源性Cdk 1，使我们能够使用大体积ATP类似物特异性地抑制Cdk 1。在同样缺乏Cdk 2的细胞中，我们发现Cdk 1活性对DNA复制起始和中心体复制至关重要。单个Cdk 2等位基因的存在使S期进展独立于Cdk 1，这表明这些激酶在S期控制中完全重叠。此外，我们发现Cdk 1抑制并没有诱导复制起点在G2期的重新许可。相反，在有丝分裂过程中的Cdk 1的抑制导致快速激活的内复制，这取决于许可抑制剂Geminin的蛋白水解。本研究证实了Cdk 1在S期调控中的重要功能，并证实了一种靶向脊椎动物细胞周期蛋白依赖性激酶的化学遗传学方法。
In vertebrates Cdk1 is required to initiate mitosis; however, any functionality of this kinase during S phase remains unclear. To investigate this, we generated chicken DT40 mutants, in which an analog-sensitive mutant cdk1 as replaces the endogenous Cdk1, allowing us to specifically inactivate Cdk1 using bulky ATP analogs. In cells that also lack Cdk2, we find that Cdk1 activity is essential for DNA replication initiation and centrosome duplication. The presence of a single Cdk2 allele renders S phase progression independent of Cdk1, which suggests a complete overlap of these kinases in S phase control. Moreover, we find that Cdk1 inhibition did not induce re-licensing of replication origins in G2 phase. Conversely, inhibition during mitosis of Cdk1 causes rapid activation of endoreplication, depending on proteolysis of the licensing inhibitor Geminin. This study demonstrates essential functions of Cdk1 in the control of S phase, and exemplifies a chemical genetics approach to target cyclin-dependent kinases in vertebrate cells.
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影响因子：
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