核酸外切酶与核酸内切酶之间的功能协作——限制酶进化的基础
Functional cooperation between exonucleases and endonucleases--basis for the evolution of restriction enzymes.
作者信息
Raghavendra Nidhanapathi K, Rao Desirazu N
机构信息
Department of Biochemistry, Indian Institute of Science, Bangalore-560012, India.
出版信息
Nucleic Acids Res. 2003 Apr 1;31(7):1888-96. doi: 10.1093/nar/gkg275.
Abstract
Many types of restriction enzymes cleave DNA away from their recognition site. Using the type III restriction enzyme, EcoP15I, which cleaves DNA 25-27 bp away from its recognition site, we provide evidence to show that an intact recognition site on the cleaved DNA sequesters the restriction enzyme and decreases the effective concentration of the enzyme. EcoP15I restriction enzyme is shown here to perform only a single round of DNA cleavage. Significantly, we show that an exonuclease activity is essential for EcoP15I restriction enzyme to perform multiple rounds of DNA cleavage. This observation may hold true for all restriction enzymes cleaving DNA sufficiently far away from their recognition site. Our results highlight the importance of functional cooperation in the modulation of enzyme activity. Based on results presented here and other data on well-characterised restriction enzymes, a functional evolutionary hierarchy of restriction enzymes is discussed.
摘要
许多类型的限制酶在远离其识别位点的地方切割DNA。使用III型限制酶EcoP15I，它在远离其识别位点25 - 27个碱基对处切割DNA，我们提供证据表明，切割后的DNA上完整的识别位点会隔离限制酶并降低酶的有效浓度。此处显示EcoP15I限制酶仅进行一轮DNA切割。值得注意的是，我们表明核酸外切酶活性对于EcoP15I限制酶进行多轮DNA切割至关重要。这一观察结果可能适用于所有在远离其识别位点处切割DNA的限制酶。我们的结果突出了功能协作在调节酶活性中的重要性。基于此处呈现的结果以及关于特征明确的限制酶的其他数据，讨论了限制酶的功能进化层次结构。
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