The structure of M.EcoKI Type I DNA methyltransferase with a DNA mimic antirestriction protein.

The structure of M.EcoKI Type I DNA methyltransferase with a DNA mimic antirestriction protein.
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DOI:
10.1093/nar/gkn988
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发表时间:
2009-02
影响因子:
14.9
通讯作者:
Dryden DT
Dryden DT
中科院分区:
生物学2区
文献类型:
--
作者:
Kennaway CK;Obarska-Kosinska A;White JH;Tuszynska I;Cooper LP;Bujnicki JM;Trinick J;Dryden DT

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I型DNA限制-修饰(R/M)系统是限制负责传播细菌对抗生素的抗性的移动的遗传元件的传播的重要试剂。EcoKI是一种来自大肠杆菌的I型R/M酶,通过甲基化和序列特异性识别起作用,导致DNA甲基化或易位,并在随机位点切割,通常相距数百个碱基对。EcoKI由一个特异性亚基、两个修饰亚基和两个DNA移位酶/核酸内切酶亚基组成,可被T7噬菌体抗限制性蛋白ocr(一种DNA模拟物)抑制。我们提出了一个三维密度图产生的负染电子显微镜和单颗粒分析的限制性复合物,M.EcoKI M2 S1甲基转移酶,结合到ocr的中央核心。我们还提出了完整的原子模型M.EcoKI与ocr及其同源DNA的复合物,给出了酶的整体钳状操作的清晰图像。该模型与40多年来发表的大量EcoKI实验数据一致。
Type-I DNA restriction–modification (R/M) systems are important agents in limiting the transmission of mobile genetic elements responsible for spreading bacterial resistance to antibiotics. EcoKI, a Type I R/M enzyme from Escherichia coli, acts by methylation- and sequence-specific recognition, leading to either methylation of DNA or translocation and cutting at a random site, often hundreds of base pairs away. Consisting of one specificity subunit, two modification subunits, and two DNA translocase/endonuclease subunits, EcoKI is inhibited by the T7 phage antirestriction protein ocr, a DNA mimic. We present a 3D density map generated by negative-stain electron microscopy and single particle analysis of the central core of the restriction complex, the M.EcoKI M2S1 methyltransferase, bound to ocr. We also present complete atomic models of M.EcoKI in complex with ocr and its cognate DNA giving a clear picture of the overall clamp-like operation of the enzyme. The model is consistent with a large body of experimental data on EcoKI published over 40 years.
DOI: 10.1093/nar/gkf518
发表时间: 2002-09-15
影响因子: 14.9
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通讯作者: Dryden, DTF
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