The bacterial DnaC helicase loader is a DnaB ring breaker.

The bacterial DnaC helicase loader is a DnaB ring breaker.
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DOI:
10.1016/j.cell.2013.03.006
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发表时间:
2013-04-11
期刊:
影响因子:
64.5
通讯作者:
Berger JM
Berger JM
中科院分区:
生物学1区
文献类型:
--
作者:
Arias-Palomo E;O'Shea VL;Hood IV;Berger JM

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专用的AAA+ ATP酶有助于将六聚环状解旋酶存款到DNA上,以促进细胞生物体中的复制。为了了解负载是如何发生的,我们使用负染电子显微镜和小角X射线散射来确定完整的E. coliDnaB·DnaC解旋酶/loader复合物。480 kDa的十二聚体形成了一个三层的组件,其中DnaC采用螺旋构型,重塑N-末端支架和C-末端DnaB的马达区,以产生解旋酶环中的明显断裂。令人惊讶的是,DnaC的AAA+折叠对于环重塑是不稳定的,因为DnaC的分离的解旋酶结合结构域既可以将DnaB加载到DNA上,又可以增加解旋酶在体外作用于底物的效率。我们的数据表明,DnaC打开DnaB的机制类似于聚合酶钳加载器,并表明细菌复制解旋酶,像他们的真核生物的同行,拥有自动调节元件,影响如何六聚体电机域加载和解开DNA。
Dedicated AAA+ ATPases help deposit hexameric ring-shaped helicases onto DNA to promote replication in cellular organisms. To understand how loading occurs, we used negative-stain electron microscopy and small-angle X-ray scattering to determine the ATP-bound structure of the intact E. coli DnaB•DnaC helicase/loader complex. The 480 kDa dodecamer forms a three-tiered assembly, in which DnaC adopts a spiral configuration that remodels N-terminal scaffolding and C-terminal motor regions of DnaB to produce a clear break in the helicase ring. Surprisingly, DnaC’s AAA+ fold is dispensable for ring remodeling, as the isolated helicase-binding domain of DnaC can both load DnaB onto DNA and increase the efficiency by which the helicase acts on substrates in vitro. Our data demonstrate that DnaC opens DnaB by a mechanism akin to that of polymerase clamp loaders, and indicate that bacterial replicative helicases, like their eukaryotic counterparts, possess auto-regulatory elements that influence how the hexameric motor domains are loaded onto and unwind DNA.
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