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骨涎蛋白限制性内切酶I DNA甲基转移酶与DNA相互作用的足迹分析。

Footprint analysis of the bsp RI DNA methyltransferase-DNA interaction.

作者信息

Finta C, Kiss A

机构信息

Institute of Biochemistry, Biological Research Center of the Hungarian Academy of Sciences, PO Box 521, Szeged 6701, Hungary.

出版信息

Nucleic Acids Res. 1997 Jul 15;25(14):2841-6. doi: 10.1093/nar/25.14.2841.

DOI:10.1093/nar/25.14.2841
PMID:9207033
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC146830/
Abstract

The interaction between the GGCC-specific Bsp RI DNA methyltransferase (M. Bsp RI) and substrate DNA was studied with footprinting techniques using a DNA fragment that was unmodified on both strands. Footprinting with DNase I revealed an approximately 14 bp protected region. Footprinting with dimethylsulfate detected major groove interactions with the guanine bases of the recognition sequence. Reaction with 1,10-phenanthroline-copper did not show protection, suggesting that minor groove interactions play little role in sequence-specific recognition by M. Bsp RI. Hydroxyl radical footprinting revealed a protected stretch of 6 nt. The hydroxyl radical footprint of M. Bsp RI differs markedly from the the footprint reported for the Hha I and Sss I methyltransferases. The pattern of protection from dimethylsulfate and hydroxyl radicals suggests that the interactions of M. Bsp RI with DNA are similar to those detected in the co-crystal structure of the Hae III methyltransferase.

摘要

使用双链均未修饰的DNA片段，通过足迹技术研究了GGCC特异性Bsp RI DNA甲基转移酶（M. Bsp RI）与底物DNA之间的相互作用。用DNase I进行足迹分析揭示了一个约14 bp的保护区域。用硫酸二甲酯进行足迹分析检测到与识别序列鸟嘌呤碱基的大沟相互作用。与1,10-菲咯啉-铜反应未显示出保护作用，这表明小沟相互作用在M. Bsp RI的序列特异性识别中作用不大。羟基自由基足迹分析揭示了一段6 nt的受保护序列。M. Bsp RI的羟基自由基足迹与报道的Hha I和Sss I甲基转移酶的足迹明显不同。硫酸二甲酯和羟基自由基的保护模式表明，M. Bsp RI与DNA的相互作用类似于在Hae III甲基转移酶的共晶体结构中检测到的相互作用。

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本文引用的文献

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A structural basis for the preferential binding of hemimethylated DNA by HhaI DNA methyltransferase.HhaI DNA甲基转移酶对半甲基化DNA优先结合的结构基础。
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