Inhibition of Ets-1 DNA binding and ternary complex formation between Ets-1, NF-κB, and DNA by a designed DNA-binding ligand
Inhibition of Ets-1 DNA binding and ternary complex formation between Ets-1, NF-κB, and DNA by a designed DNA-binding ligand
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DOI：
10.1074/jbc.274.18.12765
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发表时间：
1999-04-30
影响因子：
4.8
通讯作者：
Gottesfeld, JM
中科院分区：
文献类型：
--
作者：
Dickinson, LA;Trauger, JW;Gottesfeld, JM
序列特异性吡咯-咪唑聚酰胺可以被设计用于干扰转录因子结合和调节基因表达，无论是在体外还是在活细胞中。在人类免疫缺陷病毒1型（HnT-1）中，聚酰胺结合在TBP、ets -1和LEF-1识别位点附近，长末端重复序列抑制无细胞检测中的转录和病毒在人外周血淋巴细胞中的复制。转录因子ets - 1的DNA结合活性被结合在小凹槽中的聚酰胺特异性抑制。ets - 1是翼-螺旋-转-螺旋转录因子家族的一员，通过识别螺旋结合在主凹槽中，在主凹槽相互作用的两侧有额外的磷酸盐接触。抑制性聚酰胺可能通过占据相邻的小凹槽干扰ets - 1形成的磷酸盐接触。全长Ets-1通过与NF-kappa B的p50亚基的协同相互作用结合HIV-1增强子，Ets-1抑制聚酰胺也阻断了在HIV-1增强子上三元Ets-1 NF-kappa B DNA复合物的形成。与NF-kappa B识别位点相邻的聚酰胺结合也抑制NF-kappa B结合和三元配合物的形成。这些结果扩大了次要凹槽结合聚酰胺的应用范围，并证明这些DNA配体是DNA结合蛋白的强大抑制剂，主要使用主要凹槽接触和合作的蛋白质-DNA三元复合物。
Sequence-specific pyrrole-imidazole polyamides can be designed to interfere with transcription factor binding and to regulate gene expression, both in vitro and in living cells. Polyamides bound adjacent to the recognition sites for TBP, Ets-l, and LEF-1 in the human immunodeficiency virus, type 1 (HnT-1), long terminal repeat inhibited transcription in cell-free assays and viral rep lication in human peripheral blood lymphocytes. The DNA binding activity of the transcription factor Ets-l is specifically inhibited by a polyamide bound in the minor groove. Ets-l is a member of the winged-helix-turn-helix family of transcription factors and binds DNA through a recognition helix bound in the major groove with additional phosphate contacts on either side of this major groove interaction. The inhibitory polyamide possibly interferes with phosphate contacts made by Ets-l, by occupying the adjacent minor groove. Full-length Ets-l binds the HIV-1 enhancer through cooperative interactions with the p50 subunit of NF-kappa B, and the Ets-inhibitory polyamide also blocks formation of ternary Ets-1 NF-kappa B DNA complexes on the HIV-1 enhancer. A polyamide bound adjacent to the recognition site for NF-kappa B also inhibits NF-kappa B binding and ternary complex formation. These results broaden the application range of minor groove-binding polyamides and demonstrate that these DNA ligands are powerful inhibitors of DNA-binding proteins that predominantly use major groove contacts and of cooperative protein-DNA ternary complexes.