TY - JOUR
T1 - Targeting the DNA-binding activity of the human ERG transcription factor using new heterocyclic dithiophene diamidines
AU - Nhili, Raja
AU - Peixoto, Paul
AU - Depauw, Sabine
AU - Flajollet, Sébastien
AU - Dezitter, Xavier
AU - Munde, Manoj M.
AU - Ismail, Mohamed A.
AU - Kumar, Arvind
AU - Farahat, Abdelbasset A.
AU - Stephens, Chad E.
AU - Duterque-Coquillaud, Martine
AU - David Wilson, W.
AU - Boykin, David W.
AU - David-Cordonnier, Marie Hélène
N1 - Funding Information:
We thank the CHRU de Lille and the Région Nord/Pas-de-Calais for a PhD fellowship (to R.N.); the Institut National du Cancer (INCa) for post-doctoral fellowships (to X.D. and S.F.), and the Institut pour la Recherche sur le Cancer de Lille (IRCL) for technical expertise (to S.D.). The IMPRT-IFR114 is acknowledged for giving access to the Storm 860 equipment.
Funding Information:
The Fonds Européen de Développement Régional (FEDER, European Community) together with the Région Nord/Pas-de-Calais (to M.-H.D.-C. and M.D.-C.); the Ligue Nationale Contre le Cancer (Comité du Pas-de-Calais, Septentrion), the Association Laurette Fugain and the Association pour la Recherche sur le Cancer (to M.-H.D.-C.); NIH NIAID [064200 to W.D.W. and D.W.B.]; the CHRU de Lille and the Région Nord/Pas-de-Calais for a PhD fellowship (to R.N.); the Institut National du Cancer (INCa) for post-doctoral fellowships (to X.D. and S.F.); the Institut pour la Recherche sur le Cancer de Lille (IRCL) for technical expertise (to S.D.). Funding for open access charge: IRCL.
PY - 2013/1
Y1 - 2013/1
N2 - Direct modulation of gene expression by targeting oncogenic transcription factors is a new area of research for cancer treatment. ERG, an ETS-family transcription factor, is commonly over-expressed or translocated in leukaemia and prostate carcinoma. In this work, we selected the di-(thiophene-phenyl- amidine) compound DB1255 as an ERG/DNA binding inhibitor using a screening test of synthetic inhibitors of the ERG/DNA interaction followed by electrophoretic mobility shift assays (EMSA) validation. Spectrometry, footprint and biosensor-surface plasmon resonance analyses of the DB1255/DNA interaction evidenced sequence selectivity and groove binding as dimer. Additional EMSA evidenced the precise DNA-binding sequence required for optimal DB1255/DNA binding and thus for an efficient ERG/DNA complex inhibition. We further highlighted the structure activity relationships from comparison with derivatives. In cellulo luciferase assay confirmed this modulation both with the constructed optimal sequences and the Osteopontin promoter known to be regulated by ERG and which ERG-binding site was protected from DNaseI digestion on binding of DB1255. These data showed for the first time the ERG/DNA complex modulation, both in vitro and in cells, by a heterocyclic diamidine that specifically targets a portion of the ERG DNA recognition site.
AB - Direct modulation of gene expression by targeting oncogenic transcription factors is a new area of research for cancer treatment. ERG, an ETS-family transcription factor, is commonly over-expressed or translocated in leukaemia and prostate carcinoma. In this work, we selected the di-(thiophene-phenyl- amidine) compound DB1255 as an ERG/DNA binding inhibitor using a screening test of synthetic inhibitors of the ERG/DNA interaction followed by electrophoretic mobility shift assays (EMSA) validation. Spectrometry, footprint and biosensor-surface plasmon resonance analyses of the DB1255/DNA interaction evidenced sequence selectivity and groove binding as dimer. Additional EMSA evidenced the precise DNA-binding sequence required for optimal DB1255/DNA binding and thus for an efficient ERG/DNA complex inhibition. We further highlighted the structure activity relationships from comparison with derivatives. In cellulo luciferase assay confirmed this modulation both with the constructed optimal sequences and the Osteopontin promoter known to be regulated by ERG and which ERG-binding site was protected from DNaseI digestion on binding of DB1255. These data showed for the first time the ERG/DNA complex modulation, both in vitro and in cells, by a heterocyclic diamidine that specifically targets a portion of the ERG DNA recognition site.
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U2 - 10.1093/nar/gks971
DO - 10.1093/nar/gks971
M3 - Article
C2 - 23093599
AN - SCOPUS:84871748962
SN - 0305-1048
VL - 41
SP - 125
EP - 138
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 1
ER -
