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.
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.
Peixoto
Paul
P
INSERM U-837, Team 4-Molecular and cellular targeting for cancer treatment, Jean-Pierre Aubert Research Center, Institut de Recherches sur le Cancer de Lille, Place de Verdun, F-59045 Lille, IMPRT-IFR114, Lille, France.
Liu
Yang
Y
Depauw
Sabine
S
Hildebrand
Marie-Paule
MP
Boykin
David W
DW
Bailly
Christian
C
Wilson
W David
WD
David-Cordonnier
Marie-Hélène
MH
D020856:AT Rich Sequence; D001483:Base Sequence; D001562:Benzimidazoles / Q000494:pharmacology*; D001665:Binding Sites; D001667:Binding, Competitive; D016384:Consensus Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005663:Furans / Q000494:pharmacology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D020349:Surface Plasmon Resonance; D050818:Transcription Factor Brn-3 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D050817:Transcription Factor Pit-1 / Q000037:antagonists & inhibitors* / Q000378:metabolism
D001562:Benzimidazoles; C480104:DB293; D005663:Furans; D050818:Transcription Factor Brn-3; D050817:Transcription Factor Pit-1; D004247:DNA