Direct inhibition of the DNA-binding activity of POU transcription factors Pit-1 and Brn-3 by selective binding of a phenyl-furan-benzimidazole dication.
Direct inhibition of the DNA-binding activity of POU transcription factors Pit-1 and Brn-3 by selective binding of a phenyl-furan-benzimidazole dication.
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DOI：
10.1093/nar/gkn208
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发表时间：
2008-06
影响因子：
14.9
通讯作者：
David-Cordonnier MH
中科院分区：
文献类型：
--
作者：
Peixoto P;Liu Y;Depauw S;Hildebrand MP;Boykin DW;Bailly C;Wilson WD;David-Cordonnier MH
控制基因表达的小分子的发展可能是未来多种病理学靶向治疗方法的先锋。在为此目的开发的杂环二价阳离子中，苯基呋喃苯并咪唑二价阳离子DB 293以单体形式结合AT富集位点，以堆叠二聚体形式结合5′-ATGA序列，两者均位于小沟中。在这里，我们使用蛋白质/DNA阵列方法来评估DB293以单步竞争模式特异性抑制转录因子DNA结合的能力。DB293抑制两个POU结构域转录因子Pit-1和Brn-3，但不抑制IRF-1，尽管在所有三个共有序列中存在ATGA和AT丰富位点。EMSA、DNA酶I足迹和表面等离子体共振实验确定了DB293与共有靶点相互作用的精确结合位点、亲和力和化学计量。DB293的结合在Brn-3位点的ATGA部分以协同二聚体的形式发生，但在IRF-1序列的富含AT的位点以两个单体的形式发生。对于Pit-1位点，ATGA或富含AT的突变序列鉴定了两个位点对DB293识别的贡献。总之，DB293通过与ATGA的协同结合是两个POU结构域转录因子的强抑制剂。这些发现首次证明了杂环二价阳离子可以抑制大沟转录因子，并为这些化合物控制转录因子活性打开了大门。
The development of small molecules to control gene expression could be the spearhead of future-targeted therapeutic approaches in multiple pathologies. Among heterocyclic dications developed with this aim, a phenyl-furan-benzimidazole dication DB293 binds AT-rich sites as a monomer and 5′-ATGA sequence as a stacked dimer, both in the minor groove. Here, we used a protein/DNA array approach to evaluate the ability of DB293 to specifically inhibit transcription factors DNA-binding in a single-step, competitive mode. DB293 inhibits two POU-domain transcription factors Pit-1 and Brn-3 but not IRF-1, despite the presence of an ATGA and AT-rich sites within all three consensus sequences. EMSA, DNase I footprinting and surface-plasmon-resonance experiments determined the precise binding site, affinity and stoichiometry of DB293 interaction to the consensus targets. Binding of DB293 occurred as a cooperative dimer on the ATGA part of Brn-3 site but as two monomers on AT-rich sites of IRF-1 sequence. For Pit-1 site, ATGA or AT-rich mutated sequences identified the contribution of both sites for DB293 recognition. In conclusion, DB293 is a strong inhibitor of two POU-domain transcription factors through a cooperative binding to ATGA. These findings are the first to show that heterocyclic dications can inhibit major groove transcription factors and they open the door to the control of transcription factors activity by those compounds.
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影响因子：
10.5
作者：
Jacobson, EM;Li, P;Aggarwal, AK
通讯作者：
Aggarwal, AK
影响因子：
13.6
作者：
Fukuda, Noboru;Ueno, Takahiro;Serie, Kazuo
通讯作者：
Serie, Kazuo
影响因子：
1.3
作者：
Blagburn, BL;Drain, KL;Tidwell, RR
通讯作者：
Tidwell, RR
影响因子：
15.9
作者：
BLUME, SW;SNYDER, RC;MILLER, DM
通讯作者：
MILLER, DM
影响因子：
2.9
作者：
Bailly, C;Tardy, C;Wilson, WD
通讯作者：
Wilson, WD