Targeting the DNA-binding activity of the human ERG transcription factor using new heterocyclic dithiophene diamidines

[en] 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. <br />In this work, we selected the di-(thiophene-phenylamidine) 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 <br />further highlighted the structure activity relationshipsfrom 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.

Research center :

INSERM, U837

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Nhili, Raja

Peixoto, Paul ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > GIGA-R : Labo de recherche sur les métastases

Depauw, Sabine

Flajollet, Sebastien

Dezitter, Xavier

Munde, Manoj M.

Ismail, Mohamed

Kumar, Arvind

Farahat, Abdelbasset

Stephens, Chad

More authors (4 more)

Language :

English

Title :

Targeting the DNA-binding activity of the human ERG transcription factor using new heterocyclic dithiophene diamidines

Publication date :

2012

Journal title :

Nucleic Acids Research

ISSN :

0305-1048

eISSN :

1362-4962

Publisher :

Oxford University Press, Oxford, United Kingdom

Peer reviewed :

Peer Reviewed verified by ORBi

Funders :

FEDER, European Community, the Ligue Nationale Contre le Cancer, the Association Laurette Fugain

Commentary :

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.

Available on ORBi :

since 24 October 2012

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