Ehmann A, Chafin D, Lee K M, Hayes J J
Department of Biochemistry and Biophysics, University of Rochester, Rochester, NY 14642, USA.
Nucleic Acids Res. 1998 May 1;26(9):2086-91. doi: 10.1093/nar/26.9.2086.
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.
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。