The structural basis of Cdc7-Dbf4 kinase dependent targeting and phosphorylation of the MCM2-7 double hexamer.
The structural basis of Cdc7-Dbf4 kinase dependent targeting and phosphorylation of the MCM2-7 double hexamer.
复制标题
CDC7-Dbf4依赖于MCM2-7双六聚体的靶向和磷酸化的结构基础。
DOI：
10.1038/s41467-022-30576-1
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发表时间：
2022-05-25
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
复制叉的受控组装对于基因组稳定性至关重要。Dbf 4依赖性Cdc 7激酶（DDK）通过磷酸化Mcm 2、Mcm 4和Mcm 6的N-末端尾部处的MCM 2 -7复制解旋酶来启动复制体组装。目前，对DDK如何对接到解旋酶上以及激酶如何靶向远端Mcm亚基进行磷酸化仍然知之甚少。使用冷冻电子显微镜和生化分析，我们发现Dbf 4的HBRCT结构域与Mcm 2之间的相互作用作为锚定点，其支持DDK跨MCM 2 -7双六聚体界面的结合和Mcm 4在相对六聚体上的磷酸化。此外，DDK沿着其锚定点的旋转允许Mcm 2和Mcm 6的磷酸化。总之，我们的工作提供了基本的见解DDK结构，控制和选择性激活的MCM 2 -7解旋酶在DNA复制。重要的是，这些见解可以用于开发新型DDK抑制剂。在这里，作者描述了复制解旋酶MCM 2 -7与Dbf 4依赖性激酶（DDK）复合的多种结构。这些结构揭示了为什么激酶特异性识别MCM 2 -7双六聚体而不是单六聚体，并解释了DDK如何磷酸化位于远处的Mcm尾部。
The controlled assembly of replication forks is critical for genome stability. The Dbf4-dependent Cdc7 kinase (DDK) initiates replisome assembly by phosphorylating the MCM2-7 replicative helicase at the N-terminal tails of Mcm2, Mcm4 and Mcm6. At present, it remains poorly understood how DDK docks onto the helicase and how the kinase targets distal Mcm subunits for phosphorylation. Using cryo-electron microscopy and biochemical analysis we discovered that an interaction between the HBRCT domain of Dbf4 with Mcm2 serves as an anchoring point, which supports binding of DDK across the MCM2-7 double-hexamer interface and phosphorylation of Mcm4 on the opposite hexamer. Moreover, a rotation of DDK along its anchoring point allows phosphorylation of Mcm2 and Mcm6. In summary, our work provides fundamental insights into DDK structure, control and selective activation of the MCM2-7 helicase during DNA replication. Importantly, these insights can be exploited for development of novel DDK inhibitors. Here the authors describe multiple structures of the replicative helicase MCM2-7 in complex with Dbf4-dependent kinase (DDK). These structures reveal why the kinase specifically recognizes the MCM2-7 double-hexamer over the single-hexamer and explain how DDK can phosphorylate distantly located Mcm tails.
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影响因子：
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作者：
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DOI：
10.1107/s0907444904019158
发表时间：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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