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(1,4,7-三甲基-1,4,7-三氮杂环壬烷)铁(III)介导的DNA切割:特定蛋白质-DNA相互作用的检测

(1,4,7-trimethyl-1,4,7-triazacyclononane)iron (III)-mediated cleavage of DNA: detection of selected protein-DNA interactions.

作者信息

Ehmann A, Chafin D, Lee K M, Hayes J J

机构信息

Department of Biochemistry and Biophysics, University of Rochester, Rochester, NY 14642, USA.

出版信息

Nucleic Acids Res. 1998 May 1;26(9):2086-91. doi: 10.1093/nar/26.9.2086.

Abstract

A new reagent for the oxidative cleavage of DNA, (1,4,7-trimethyl-1, 4,7-triazacyclononane)iron(III) chloride was recently introduced. We have determined the utility of this reagent for detecting protein-DNA interactions within two types of complexes. Interestingly, we find that the rates of DNA cleavage by this reagent are differentially affected by the two classes of protein-DNA interactons studied. We find that the rate of DNA cleavage by this reagent is relatively unaffected by the non-sequence-specific histone-DNA interactions within a nucleosome complex. Conversely, a clear footprint pattern is obtained with two different DNA sequence-specific protein-DNA complexes. The results suggest that (1,4,7-trimethyl-1,4,7-triazacyclononane)iron(III) chloride will be a useful reagent to probe trans -acting-factor-DNA interactions within a chromatin environment. Differences between these two types of protein-DNA interactions, which might account for this observation, are discussed.

摘要

最近引入了一种用于DNA氧化切割的新试剂,即(1,4,7-三甲基-1,4,7-三氮杂环壬烷)氯化铁(III)。我们已经确定了该试剂在检测两种复合物中蛋白质-DNA相互作用方面的效用。有趣的是,我们发现该试剂切割DNA的速率受到所研究的两类蛋白质-DNA相互作用的不同影响。我们发现该试剂切割DNA的速率相对不受核小体复合物中非序列特异性组蛋白-DNA相互作用的影响。相反,使用两种不同的DNA序列特异性蛋白质-DNA复合物可获得清晰的足迹模式。结果表明,(1,4,7-三甲基-1,4,7-三氮杂环壬烷)氯化铁(III)将是在染色质环境中探测反式作用因子-DNA相互作用的有用试剂。本文讨论了这两种蛋白质-DNA相互作用之间的差异,这些差异可能解释了这一观察结果。

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本文引用的文献

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Chemical probes of DNA structure in chromatin.
Chem Biol. 1995 Mar;2(3):127-35. doi: 10.1016/1074-5521(95)90066-7.

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