Single strand transposition at the host replication fork.
Single strand transposition at the host replication fork.
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DOI：
10.1093/nar/gkw661
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发表时间：
2016-09-19
影响因子：
14.9
通讯作者：
Ton-Hoang B
中科院分区：
文献类型：
--
作者：
Lavatine L;He S;Caumont-Sarcos A;Guynet C;Marty B;Chandler M;Ton-Hoang B
IS200/IS605 插入序列家族的成员与经典 IS 的根本区别在于其特定的单链 (ss) 转座机制，该机制由 Y1 转座酶 TnpA（一种识别和处理 ss DNA 底物的小型 HuH 酶）精心策划。转座通过“剥离和粘贴”途径发生，该途径由两个步骤组成：精确切除顶链作为环状 ss DNA 中间体；并随后整合到特定的 ssDNA 靶标中。通过计算机高通量分析实验显示或暗示家族成员的转座与复制叉的滞后链模板紧密耦合。在这项研究中，我们研究了复制叉靶向相关的因素，并分析了转座酶的 DNA 结合特性，转座酶可以帮助复制叉上 ss DNA 底物的定位。我们证明 TnpA 与 β 滑动钳 DnaN 相互作用并识别模仿复制叉结构的 DNA。我们还表明，dsDNA 可以促进 TnpA 靶向 ssDNA 底物。我们在体外分析了 Ssb 和 RecA 蛋白对 TnpA 活性的影响，结果表明，虽然 RecA 没有表现出显着的影响，但 Ssb 抑制整合。最后，我们讨论了在复制叉处整合到 ssDNA 中的方式。
Members of the IS200/IS605 insertion sequence family differ fundamentally from classical IS essentially by their specific single-strand (ss) transposition mechanism, orchestrated by the Y1 transposase, TnpA, a small HuH enzyme which recognizes and processes ss DNA substrates. Transposition occurs by the ‘peel and paste’ pathway composed of two steps: precise excision of the top strand as a circular ss DNA intermediate; and subsequent integration into a specific ssDNA target. Transposition of family members was experimentally shown or suggested by in silico high-throughput analysis to be intimately coupled to the lagging strand template of the replication fork. In this study, we investigated factors involved in replication fork targeting and analysed DNA-binding properties of the transposase which can assist localization of ss DNA substrates on the replication fork. We showed that TnpA interacts with the β sliding clamp, DnaN and recognizes DNA which mimics replication fork structures. We also showed that dsDNA can facilitate TnpA targeting ssDNA substrates. We analysed the effect of Ssb and RecA proteins on TnpA activity in vitro and showed that while RecA does not show a notable effect, Ssb inhibits integration. Finally we discuss the way(s) in which integration may be directed into ssDNA at the replication fork.
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影响因子：
14.9
作者：
He S;Guynet C;Siguier P;Hickman AB;Dyda F;Chandler M;Ton-Hoang B
通讯作者：
Ton-Hoang B
影响因子：
5.6
作者：
GALAS, DJ;CHANDLER, M
通讯作者：
CHANDLER, M
影响因子：
11.4
作者：
Bouvier, M;Demarre, G;Mazel, D
通讯作者：
Mazel, D
影响因子：
4.5
作者：
Ah-Seng, Yoan;Rech, Jerome;Bouet, Jean-Yves
通讯作者：
Bouet, Jean-Yves
影响因子：
16.8
作者：
Georgescu, Roxana E.;Kurth, Isabel;O'Donnell, Mike E.
通讯作者：
O'Donnell, Mike E.