Binding of distamycin and chromomycin to human immunodeficiency type 1 virus DNA: a non-radiactive automated footprinting study

Giordana Feriotto, Carlo Mischiati, Nicoletta Bianchi, Marco Passadore, Roberto Gambari
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引用次数: 11

Abstract

Sequence-selectivity of DNA-binding drugs was recently reported in a number of studies employing footprinting and gel retardation approaches. In this paper we studied sequence-selectivity of the binding of chromomycin and distamycin to DNA by performing DNase I footprinting and analysis of the cleaved fragments by the Pharmacia ALFTM DNA Sequencing System. As a model system we employed the long terminal repeat of the human immunodeficiency type 1 virus. The main conclusion of our experiments is that automated analysis of DNase I footprinting is a fast and reliable technique to study drugs-DNA interactions. The results obtained suggest that distamycin and chromomycin differentially interact with the long terminal repeat of the human immunodeficiency type 1 virus; this differential binding depends upon the DNA sequences recognized. The data presented are consistent with a preferential binding of distamycin to DNA sequences of the binding sites of nuclear factor kappa B and transcription factor IID. By contrast, distamycin exhibits only weak binding to DNA sequences recognized by the promoter-specific transcription factor Sp1. Unlike distamycin, chromomycin preferentially interacts with the binding sites of the promoter-specific transcription factor Sp1.

二霉素和色霉素与人类免疫缺陷1型病毒DNA的结合:一项非放射性自动足迹研究
DNA结合药物的序列选择性最近在许多采用足迹法和凝胶阻滞法的研究中被报道。在本文中,我们通过DNA酶I足迹和Pharmacia ALFTM DNA测序系统对切割片段的分析,研究了色霉素和远端霉素与DNA结合的序列选择性。作为模型系统,我们采用了人类免疫缺陷1型病毒的长末端重复序列。我们实验的主要结论是,DNA酶I足迹的自动分析是研究药物与DNA相互作用的一种快速可靠的技术。所获得的结果表明,远端霉素和色霉素与人类免疫缺陷1型病毒的长末端重复序列有不同的相互作用;这种差异结合取决于所识别的DNA序列。所提供的数据与远端霉素与核因子κB和转录因子IID结合位点的DNA序列的优先结合一致。相反,远端霉素仅表现出与启动子特异性转录因子Sp1识别的DNA序列的弱结合。与远霉素不同,色霉素优先与启动子特异性转录因子Sp1的结合位点相互作用。
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