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EcoPI限制酶切割DNA需要ATP水解。

ATP hydrolysis is required for DNA cleavage by EcoPI restriction enzyme.

作者信息

Saha S, Rao D N

机构信息

Department of Biochemistry, Indian Institute of Science, Bangalore.

出版信息

J Mol Biol. 1995 Apr 7;247(4):559-67. doi: 10.1016/s0022-2836(05)80137-8.

DOI:10.1016/s0022-2836(05)80137-8
PMID:7723013
Abstract

The type III restriction endonuclease EcoPI, coded by bacteriophage P1, cleaves unmodified DNA in the presence of ATP and magnesium ions. We show that purified EcoPI restriction enzyme fails to cleave DNA in the presence of non-hydrolyzable ATP analogs. More importantly, this study demonstrates that EcoPI restriction enzyme has an inherent ATPase activity, and ATP hydrolysis is necessary for DNA cleavage. Furthermore, we show that the progress curve of the reaction with EcoPI restriction enzyme exhibits a lag which is dependent on the enzyme concentration. Kinetic analysis of the progress curves of the reaction suggest slow transitions that can occur during the reaction, characteristic of hysteretic enzymes. The role of ATP in the cleavage mechanism of type III restriction enzymes is discussed.

摘要

由噬菌体P1编码的III型限制性内切酶EcoPI，在ATP和镁离子存在的情况下切割未修饰的DNA。我们发现，纯化的EcoPI限制酶在存在不可水解的ATP类似物时无法切割DNA。更重要的是，本研究表明EcoPI限制酶具有内在的ATP酶活性，并且ATP水解是DNA切割所必需的。此外，我们表明，EcoPI限制酶反应的进程曲线呈现出一个依赖于酶浓度的滞后现象。对反应进程曲线的动力学分析表明，反应过程中可能会发生缓慢的转变，这是滞后酶的特征。本文讨论了ATP在III型限制酶切割机制中的作用。

相似文献

1
ATP hydrolysis is required for DNA cleavage by EcoPI restriction enzyme.EcoPI限制酶切割DNA需要ATP水解。
J Mol Biol. 1995 Apr 7;247(4):559-67. doi: 10.1016/s0022-2836(05)80137-8.
2
Mutations in the Res subunit of the EcoPI restriction enzyme that affect ATP-dependent reactions.影响ATP依赖性反应的EcoPI限制酶Res亚基中的突变。
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3
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S-adenosyl homocysteine and DNA ends stimulate promiscuous nuclease activities in the Type III restriction endonuclease EcoPI.S-腺苷同型半胱氨酸和DNA末端刺激III型限制性内切酶EcoPI中的混杂核酸酶活性。
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Functional analysis of conserved motifs in type III restriction-modification enzymes.III型限制修饰酶中保守基序的功能分析
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DNA recognition by the EcoP15I and EcoPI modification methyltransferases.EcoP15I和EcoPI甲基化修饰转移酶对DNA的识别
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Type III restriction endonucleases are heterotrimeric: comprising one helicase-nuclease subunit and a dimeric methyltransferase that binds only one specific DNA.III 型限制内切酶是异源三聚体：由一个解旋酶-核酸酶亚基和一个二聚体甲基转移酶组成，后者仅与一种特定的 DNA 结合。
Nucleic Acids Res. 2014 Apr;42(8):5139-50. doi: 10.1093/nar/gku122. Epub 2014 Feb 7.

引用本文的文献

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DNA cleavage site selection by Type III restriction enzymes provides evidence for head-on protein collisions following 1D bidirectional motion.III 型限制酶对 DNA 切割位点的选择为沿 1D 双向运动的蛋白迎面相撞提供了证据。
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