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Distamycin A and tallimustine inhibit TBP binding and basal in vitro transcription.

机译:Distamycin A和图莫司汀抑制TBP结合和基础体外转录。

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摘要

The antibiotic distamycin A is a DNA minor groove binding drug (MGB) that recognizes a stretch of at least four ATs. The alkylating benzoyl mustard derivative tallimustine (FCE 24517) has powerful anti-tumor activity. Using the electrophoretic mobility shift assay (EMSA) we determined that both compounds can prevent binding of TBP and, with 10-fold higher concentration, TBP-TFIIA (DA) and TBP-TFIIA-TFIIB (DAB) to a TATA box. Once formed, the DA and DAB complexes are more resistant to MGB challenge. Both drugs can inhibit basal in vitro transcription of a minimal TATA-containing promoter and similar concentrations are necessary for binding and transcriptional inhibition. Tallimustine shows strong selectivity by decreasing only correctly initiated transcripts. Even at high doses (20 microM), however, they cannot disturb a competent pre-initiation complex or Pol II progression. This functional in vitro model will provide a way to investigate the activity of sequence-specific DNA binding drugs with potential anti-viral and anti-tumour activity and to develop novel more selective compounds.
机译:抗生素双歧霉素A是可识别至少四个AT的DNA小沟结合药物(MGB)。烷基化的苯甲酰芥子碱衍生物妥西莫司汀(FCE 24517)具有强大的抗肿瘤活性。使用电泳迁移率变动分析(EMSA),我们确定了这两种化合物都可以防止TBP结合,并且以10倍高的浓度将TBP-TFIIA(DA)和TBP-TFIIA-TFIIB(DAB)结合到TATA盒上。一旦形成,DA和DAB复合物就更能抵抗MGB攻击。两种药物都可以抑制包含TATA的最小启动子的基础体外转录,并且相似的浓度对于结合和转录抑制是必需的。塔利莫司汀通过仅减少正确起始的转录本而显示出强选择性。但是,即使在高剂量(20 microM)下,它们也不会干扰有效的预起始复合物或Pol II进展。这种功能性体外模型将提供一种方法,以研究具有潜在抗病毒和抗肿瘤活性的序列特异性DNA结合药物的活性,并开发出更具选择性的新型化合物。

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