Pseudocomplementary PNAs as selective modifiers of protein activity on duplex DNA: the case of type IIs restriction enzymes.
Pseudocomplementary PNAs as selective modifiers of protein activity on duplex DNA: the case of type IIs restriction enzymes.
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伪互补 PNA 作为双链 DNA 上蛋白质活性的选择性修饰剂：II 型限制酶的情况。
DOI：
10.1093/nar/gkg450
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发表时间：
2003
影响因子：
14.9
通讯作者：
Frank-Kamenetskii,MaximD
中科院分区：
文献类型：
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作者：
Protozanova,Ekaterina;Demidov,VadimV;Nielsen,PeterE;Frank-Kamenetskii,MaximD
本研究评估了假互补肽核酸（pcPNAs）对双链DNA （dsDNA）酶活性序列特异性修饰的潜力。为此，我们分析了pcPNA-dsDNA复合物位点选择性干扰四种ii型限制性内切酶的能力。我们发现pcPNA-dsDNA复合物对不同的同分异构体内切酶（PleI， MlyI和N.BstNBI）的切割/缺口活性表现出不同程度的DNA保护，这取决于它们的类型和PNA结合位点和酶识别/切割位点的相互排列。我们还发现，pcPNA靶向位于dsDNA上的PleI或BbsI识别位点，在某些情况下会产生这些DNA切割器的切割活性。同时，PleI同分异构体MlyI内切酶不表现出任何DNA切割活性，被附近的pcPNA结合完全阻断。我们的研究结果对有效的pcPNA干扰DNA加工蛋白的性能具有普遍意义，因此对pcPNA的未来应用具有重要意义。
This study evaluates the potential of pseudocomplementary peptide nucleic acids (pcPNAs) for sequence‐specific modification of enzyme activity towards double‐stranded DNA (dsDNA). To this end, we analyze the ability of pcPNA–dsDNA complexes to site‐selectively interfere with the action of four type IIs restriction enzymes. We have found that pcPNA–dsDNA complexes exhibit a different degree of DNA protection against cleaving/nicking activity of various isoschizomeric endonucleases under investigation (PleI, MlyI and N.BstNBI) depending on their type and mutual arrangement of PNA‐binding and enzyme recognition/cleavage sites. We have also found that the pcPNA targeting to closely located PleI or BbsI recognition sites on dsDNA generates in some cases the nicking activity of these DNA cutters. At the same time, MlyI endonuclease, a PleI isoschizomer, does not exhibit any DNA nicking/cleavage activity, being completely blocked by the nearby pcPNA binding. Our results have general implications for effective pcPNA interference with the performance of DNA‐processing proteins, thus being important for prospective applications of pcPNAs.