Crystal structure of E. coli RecE protein reveals a toroidal tetramer for processing double-stranded DNA breaks.
Crystal structure of E. coli RecE protein reveals a toroidal tetramer for processing double-stranded DNA breaks.
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DOI：
10.1016/j.str.2009.03.008
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发表时间：
2009-05-13
期刊：
影响因子：
--
通讯作者：
Bell CE
中科院分区：
文献类型：
--
作者：
Zhang J;Xing X;Herr AB;Bell CE
大肠杆菌RecE蛋白是经典RecET重组系统的一部分，该系统最近被用于强大的基因工程新方法。RecE结合到游离的dsDNA末端，并逐渐消化5 ‘末端的链，形成5 ’ -单核苷酸和3 ' -悬垂物，这是RecT促进单链退火的底物。在这里，我们以2.8 Å的分辨率报道了RecE的c端核酸酶结构域的晶体结构。RecE形成一个环形四聚体，其中心有一个足够宽的锥形通道，可以在一端结合dsDNA，但在另一端被蛋白质的c端片段部分堵塞。四个狭窄的隧道，在四聚体的每个亚基内，从中央通道通向四个活性位点，它们距离通道约15 Å。结合突变研究，该结构表明dsDNA通过中心通道的开放端进入，5 ‘端链通过隧道进入四个活性位点之一，3 ’端链通过四聚体后部通道的堵塞端。
E. coli RecE protein is part of the classical RecET recombination system that has recently been employed in powerful new methods for genetic engineering. RecE binds to free dsDNA ends and processively digests the 5′-ended strand to form 5′-mononucleotides and a 3′-overhang that is a substrate for single strand annealing promoted by RecT. Here, we report the crystal structure of the C-terminal nuclease domain of RecE at 2.8 Å resolution. RecE forms a toroidal tetramer with a central tapered channel that is wide enough to bind dsDNA at one end, but is partially plugged at the other end by the C-terminal segment of the protein. Four narrow tunnels, one within each subunit of the tetramer, lead from the central channel to the four active sites, which lie about 15 Å from the channel. The structure, combined with mutational studies, suggests a mechanism in which dsDNA enters through the open end of the central channel, the 5′-ended strand passes through a tunnel to access one of the four active sites, and the 3′-ended strand passes through the plugged end of the channel at the back of the tetramer.
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DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
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通讯作者：
Cowtan, K
影响因子：
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作者：
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通讯作者：
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DOI：
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发表时间：
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期刊：
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影响因子：
--
作者：
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通讯作者：
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DOI：
10.1107/s0907444902016657
发表时间：
2002-11-01
影响因子：
2.2
作者：
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通讯作者：
Terwilliger, TC
影响因子：
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作者：
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通讯作者：
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