ATP-dependent conformational change in ABC-ATPase RecF serves as a switch in DNA repair.
ATP-dependent conformational change in ABC-ATPase RecF serves as a switch in DNA repair.
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ABC-ATPase RecF 中 ATP 依赖性构象变化充当 DNA 修复的开关。
DOI：
10.1038/s41598-018-20557-0
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发表时间：
2018-02-01
影响因子：
4.6
通讯作者：
Yan XX
中科院分区：
文献类型：
--
作者：
Tang Q;Liu YP;Shan HH;Tian LF;Zhang JZ;Yan XX
RecF是RecF通路的主要成员。它与RecO和RecR相互作用，通过将RecA重组酶装载在单链DNA上并置换单链DNA结合蛋白来启动同源重组。作为ATP结合盒ATP酶，RecF表现出ATP依赖的二聚化和与Rad 50和SMC蛋白的结构同源性。然而，RecF ATP依赖性二聚化的机制和作用模式仍不清楚。在此，我们确定了TTERecF，TTERecF-ATP和TTERecF-ATP三种晶体结构，揭示了一种新的ATP驱动的RecF二聚化。RecF在其二聚体界面上含有一个带正电荷的隧道，这对ATP结合至关重要。我们的结构和生化数据表明，步行者A基序作为一个开关，并在ATP结合和RecF二聚化中发挥关键作用。此外，生物层干涉法测定结果表明，TTERecF与ATP相互作用，形成二聚体，显示出比TTERecF单体更高的DNA亲和力。总之，我们的研究结果为理解RecF与ATP结合的过程和ATP依赖的RecF二聚化的功能机制提供了坚实的结构基础。
RecF is a principal member of the RecF pathway. It interacts with RecO and RecR to initiate homologous recombination by loading RecA recombinases on single-stranded DNA and displacing single-stranded DNA-binding proteins. As an ATP-binding cassette ATPase, RecF exhibits ATP-dependent dimerization and structural homology with Rad50 and SMC proteins. However, the mechanism and action pattern of RecF ATP-dependent dimerization remains unclear. Here, We determined three crystal structures of TTERecF, TTERecF-ATP and TTERecF-ATPɤS fromThermoanaerobacter tengcongensisthat reveal a novel ATP-driven RecF dimerization. RecF contains a positively charged tunnel on its dimer interface that is essential to ATP binding. Our structural and biochemical data indicate that the Walker A motif serves as a switch and plays a key role in ATP binding and RecF dimerization. Furthermore, Biolayer interferometry assay results showed that the TTERecF interacted with ATP and formed a dimer, displaying a higher affinity for DNA than that of the TTERecF monomer. Overall, our results provide a solid structural basis for understanding the process of RecF binding with ATP and the functional mechanism of ATP-dependent RecF dimerization.
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影响因子：
14.9
作者：
Radzimanowski J;Dehez F;Round A;Bidon-Chanal A;McSweeney S;Timmins J
通讯作者：
Timmins J
影响因子：
3.6
作者：
Kidane, D;Sanchez, H;Graumann, PL
通讯作者：
Graumann, PL
影响因子：
5.6
作者：
Moncalian, G;Lengsfeld, B;Paull, TT
通讯作者：
Paull, TT
影响因子：
5.6
作者：
Shan, Q;Bork, JM;Cox, MM
通讯作者：
Cox, MM
影响因子：
3.2
作者：
Lenhart, Justin S.;Brandes, Eileen R.;Simmons, Lyle A.
通讯作者：
Simmons, Lyle A.