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多胺和碱性蛋白质对限制性核酸内切酶切割DNA的影响。

Effect of polyamines and basic proteins on cleavage of DNA by restriction endonucleases.

作者信息

Pingoud A, Urbanke C, Alves J, Ehbrecht H J, Zabeau M, Gualerzi C

出版信息

Biochemistry. 1984 Nov 20;23(24):5697-703. doi: 10.1021/bi00319a006.

Abstract

We have investigated the effect of the polyamines spermine, spermidine, and putrescine and the prokaryotic histone-like proteins NS1 and NS2 on the restriction endonuclease EcoRI catalyzed cleavage of plasmid and bacteriophage DNAs. At low concentrations of spermine and spermidine, the rate of DNA cleavage by EcoRI is increased, while high concentrations of spermine as well as of spermidine are inhibitory. These phenomena are also observed with other restriction endonucleases. They are, therefore, probably due to the interaction of the polyamines with the DNA. Putrescine does not have such an effect within the concentration range investigated. Remarkably, low concentrations of spermine and spermidine very efficiently suppress EcoRI activity. An inhibition of the EcoRI-catalyzed cleavage of DNA is also observed with NS1 and NS2, an effect that can be mimicked with other basic proteins that interact with DNA. The results are discussed in terms of the mechanism of restriction in vivo.

摘要

我们研究了多胺精胺、亚精胺和腐胺以及原核生物类组蛋白NS1和NS2对限制性内切酶EcoRI催化的质粒和噬菌体DNA切割的影响。在精胺和亚精胺浓度较低时,EcoRI切割DNA的速率增加,而高浓度的精胺和亚精胺则具有抑制作用。其他限制性内切酶也观察到了这些现象。因此,这些现象可能是由于多胺与DNA的相互作用所致。在所研究的浓度范围内,腐胺没有这种作用。值得注意的是,低浓度的精胺和亚精胺能非常有效地抑制EcoRI活性。NS1和NS2也观察到对EcoRI催化的DNA切割有抑制作用,其他与DNA相互作用的碱性蛋白也能模拟这种作用。根据体内限制机制对结果进行了讨论。

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