IS200/IS605 family single-strand transposition: mechanism of IS608 strand transfer.
IS200/IS605 family single-strand transposition: mechanism of IS608 strand transfer.
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DOI：
10.1093/nar/gkt014
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发表时间：
2013-03-01
影响因子：
14.9
通讯作者：
Ton-Hoang B
中科院分区：
文献类型：
--
作者：
He S;Guynet C;Siguier P;Hickman AB;Dyda F;Chandler M;Ton-Hoang B
IS200/IS605家族成员IS608的转座酶TnpA催化单链DNA转座，并且是具有杂合催化位点的二聚体，所述杂合催化位点由来自一个单体的HUH基序和存在于来自另一个单体的α-螺旋（α D）中的催化Y127（反式构型）组成。α D通过柔性环连接到主体。虽然导致切除的转座中间体的反应是很好的特点，很少有人知道的动力学行为的转座体驱动这一进程。我们提供的证据强烈支持链转移模型，涉及旋转的α D螺旋从反式顺式构型（HUH和Y残基来自同一单体）。对TnpA异二聚体的研究表明TnpA以反式构型切割DNA，并且与5 ′-磷酸盐连接的催化酪氨酸交换位置以允许以顺式构型重新连接切割的链（链转移）。它们进一步暗示，在切除转座子连接后，TnpA应在整合所需的切割之前重置为反式构型。分析还表明，这种机制在IS200/IS605家族成员中是保守的。
Transposase, TnpA, of the IS200/IS605 family member IS608, catalyses single-strand DNA transposition and is dimeric with hybrid catalytic sites composed of an HUH motif from one monomer and a catalytic Y127 present in an α-helix (αD) from the other (trans configuration). αD is attached to the main body by a flexible loop. Although the reactions leading to excision of a transposition intermediate are well characterized, little is known about the dynamic behaviour of the transpososome that drives this process. We provide evidence strongly supporting a strand transfer model involving rotation of both αD helices from the trans to the cis configuration (HUH and Y residues from the same monomer). Studies with TnpA heterodimers suggest that TnpA cleaves DNA in the trans configuration, and that the catalytic tyrosines linked to the 5′-phosphates exchange positions to allow rejoining of the cleaved strands (strand transfer) in the cis configuration. They further imply that, after excision of the transposon junction, TnpA should be reset to a trans configuration before the cleavage required for integration. Analysis also suggests that this mechanism is conserved among members of the IS200/IS605 family.
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影响因子：
5.6
作者：
GALAS, DJ;CHANDLER, M
通讯作者：
CHANDLER, M
影响因子：
14.9
作者：
He S;Hickman AB;Dyda F;Johnson NP;Chandler M;Ton-Hoang B
通讯作者：
Ton-Hoang B
影响因子：
16
作者：
Guynet, Catherine;Achard, Adeline;Chandler, Michael
通讯作者：
Chandler, Michael
影响因子：
64.5
作者：
Barabas, Orsolya;Ronning, Donald R.;Dyda, Fred
通讯作者：
Dyda, Fred
影响因子：
5.6
作者：
CHANDLER, M;GALAS, DJ
通讯作者：
GALAS, DJ