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首页> 外文期刊>Journal of Molecular Biology >The Escherichia coli dam DNA methyltransferase modifies DNA in a highly processive reaction.
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The Escherichia coli dam DNA methyltransferase modifies DNA in a highly processive reaction.

机译：大肠杆菌大坝DNA甲基转移酶以高度加工性反应修饰DNA。

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摘要

The Escherichia coli dam adenine-N6 methyltransferase modifies DNA at GATC sequences. It is involved in post-replicative mismatch repair, control of DNA replication and gene regulation. We show that E. coli dam acts as a functional monomer and methylates only one strand of the DNA in each binding event. The preferred way of ternary complex assembly is that the enzyme first binds to DNA and then to S-adenosylmethionine. The enzyme methylates an oligonucleotide containing two dam sites and a 879 bp PCR product with four sites in a fully processive reaction. On lambda-DNA comprising 48,502 bp and 116 dam sites, E. coli dam scans 3000 dam sites per binding event in a random walk, that on average leads to a processive methylation of 55 sites. Processive methylation of DNA considerably accelerates DNA methylation. The highly processive mechanism of E. coli dam could explain why small amounts of E. coli dam are able to maintain the methylation state of dam sites during DNA replication. Furthermore, our data support the general rule that solitary DNA methyltransferase modify DNA processively whereas methyltransferases belonging to a restriction-modification system show a distributive mechanism, because processive methylation of DNA would interfere with the biological function of restriction-modification systems.
机译：大肠杆菌大坝腺嘌呤-N6甲基转移酶修饰GATC序列的DNA。它参与复制后错配修复，DNA复制控制和基因调控。我们显示大肠杆菌大坝充当功能性单体，并且在每次结合事件中仅甲基化DNA的一条链。三元复合物组装的优选方式是该酶首先结合DNA，然后结合S-腺苷甲硫氨酸。该酶在完全进行的反应中甲基化了一个寡核苷酸，该寡核苷酸包含两个dam位点和一个带有四个位点的879 bp PCR产物。在包含48,502 bp的Lambda DNA和116个dam位点上，大肠杆菌dam随机结合每个结合事件扫描3000个dam位点，平均导致55个位点的进行性甲基化。 DNA的进行性甲基化大大加速了DNA甲基化。 E. coli dam的高度加工机制可以解释为什么在DNA复制过程中少量E. coli dam能够维持Dam部位的甲基化状态。此外，我们的数据支持一般规则，即单独的DNA甲基转移酶会进行DNA修饰，而属于限制性修饰系统的甲基转移酶则显示出分布机制，因为DNA的甲基化会干扰限制性修饰系统的生物学功能。

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