PcrA-mediated disruption of RecA nucleoprotein filaments--essential role of the ATPase activity of RecA.

PcrA-mediated disruption of RecA nucleoprotein filaments--essential role of the ATPase activity of RecA.
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DOI:
10.1093/nar/gks641
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发表时间:
2012-09-01
影响因子:
14.9
通讯作者:
Anand SP
Anand SP
中科院分区:
生物学2区
文献类型:
--
作者:
Fagerburg MV;Schauer GD;Thickman KR;Bianco PR;Khan SA;Leuba SH;Anand SP

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必需的DNA解旋酶PcrA通过取代DNA中的重组酶RecA来调节重组。RecA的核苷酸结合状态决定了其核蛋白细丝的稳定性。使用单分子荧光方法,我们证明了通过移位PcrA来置换RecA需要重组酶的ATPase活性。我们还表明,在聚合的RecA细丝和移位的PcrA之间的“迎头碰撞”中,RecA K72R ATPase突变体,而不是野生型RecA,阻止了解旋酶的易位。我们的发现表明,PcrA的易位不足以取代DNA中的RecA,并且在解旋酶介导的细丝破坏中,RecA的ATPase活性起着至关重要的作用。
The essential DNA helicase, PcrA, regulates recombination by displacing the recombinase RecA from the DNA. The nucleotide-bound state of RecA determines the stability of its nucleoprotein filaments. Using single-molecule fluorescence approaches, we demonstrate that RecA displacement by a translocating PcrA requires the ATPase activity of the recombinase. We also show that in a ‘head-on collision’ between a polymerizing RecA filament and a translocating PcrA, the RecA K72R ATPase mutant, but not wild-type RecA, arrests helicase translocation. Our findings demonstrate that translocation of PcrA is not sufficient to displace RecA from the DNA and assigns an essential role for the ATPase activity of RecA in helicase-mediated disruption of its filaments.
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