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Research Articles: Therapeutics, Targets, and Development

Inhibition of DNA binding of the NF-Y transcription factor by the pyrrolobenzodiazepine-polyamide conjugate GWL-78

¹ Cancer Research UK Drug-DNA Interactions Research Group, UCL Cancer Institute, University College London; ² Cancer Research UK Gene Targeted Drug Design Research Group, School of Pharmacy, University of London, London, United Kingdom; and ³ Dipartimento di Scienze Biomolecolari e Biotecnologie, Universita degli Studi di Milano, Milan, Italy

Requests for reprints: Daniel Hochhauser, UCL Cancer Institute, 72 Huntley Street, London WCIE 6RT, United Kingdom. Phone: 44-20-7679-6006. E-mail: d.hochhauser{at}ucl.ac.uk

Abstract

Many genes involved in cell cycle control have promoters that bind the heterotrimeric transcription factor NF-Y. Several minor-groove binding drugs have been shown to block interactions of transcription factors with cognate DNA-binding sequences. We showed previously that noncovalent minor-groove binding agents block interactions of NF-Y with the promoter of topoisomerase II (topo II). In this study, we investigated the ability of GWL-78, a pyrrolobenzodiazepine-poly(N-methylpyrrole) conjugate, to inhibit the binding of NF-Y to DNA. Electrophoretic mobility shift assays showed that GWL-78 could displace NF-Y bound to several CCAAT motifs within promoters of genes involved in cell cycle progression. DNase I footprinting of the topo II promoter confirmed binding of GWL-78 to AT-rich sequences corresponding to the preferred binding site of NF-Y. Incubation with GWL-78 resulted in displacement of NF-Y binding to DNA. Chromatin immunoprecipitation assays on the topo II promoter showed that GWL-78 was able to enter the nucleus and interact with specific DNA sequences. Treatment of NIH3T3 cells with GWL-78 resulted in a block of cell cycle progression, which did not involve activation of p53. Thus, agents such as GWL-78 may be useful in modulating transcription and blocking cellular proliferation. [Mol Cancer Ther 2008;7(5):1319–28]

Grant support: MRC Career Establishment Grant G0000168 (D. Hochhauser) and Cancer Research UK programme C2559/A3083 (J.A. Hartley and D. Hochhauser).

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

⁴ Ceribelli et al., submitted for publication.

Received 7/16/07; revised 2/ 5/08; accepted 3/18/08.