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Molecular and Cellular Biology, November 2002, p. 7769-7779, Vol. 22, No. 22
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.22.7769-7779.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Oct-1 Potentiates CREB-Driven Cyclin D1 Promoter Activation via a Phospho-CREB- and CREB Binding Protein-Independent Mechanism

Séverine Boulon, Jean-Christophe Dantonel, Virginie Binet, Annick Vié, Jean-Marie Blanchard, Robert A. Hipskind, and Alexandre Philips*

Institut de Génétique Moléculaire, CNRS, UMR 5535, IFR24, 34293 Montpellier Cedex 5, France

Received 27 March 2002/ Returned for modification 19 June 2002/ Accepted 9 August 2002

Cyclin D1, the regulatory subunit for mid-G1 cyclin-dependent kinases, controls the expression of numerous cell cycle genes. A cyclic AMP-responsive element (CRE), located upstream of the cyclin D1 mRNA start site, integrates mitogenic signals that target the CRE-binding factor CREB, which can recruit the transcriptional coactivator CREB-binding protein (CBP). We describe an alternative mechanism for CREB-driven cyclin D1 induction that involves the ubiquitous POU domain protein Oct-1. In the breast cancer cell line MCF-7, overexpression of Oct-1 or its POU domain strongly increases transcriptional activation of cyclin D1 and GAL4 reporter genes that is specifically dependent upon CREB but independent of Oct-1 DNA binding. Gel retardation and chromatin immunoprecipitation assays confirm that POU forms a complex with CREB bound to the cyclin D1 CRE. In solution, CREB interaction with POU requires the CREB Q2 domain and, notably, occurs with CREB that is not phosphorylated on Ser 133. Accordingly, Oct-1 also potently enhances transcriptional activation mediated by a Ser133Ala CREB mutant. Oct-1/CREB synergy is not diminished by the adenovirus E1A 12S protein, a repressor of CBP coactivator function. In contrast, E1A strongly represses CBP-enhanced transactivation by CREB phosphorylated on Ser 133. Our observation that Oct-1 potentiates CREB-dependent cyclin D1 transcriptional activity independently of Ser 133 phosphorylation and E1A-sensitive coactivator function offers a new paradigm for the regulation of cyclin D1 induction by proliferative signals.

* Corresponding author. Mailing address: Institut de Génétique Moléculaire, CNRS, UMR 5535, IFR24, 1919 Route de Mende, 34293 Montpellier Cedex 5, France. Phone: 33 4 67613650. Fax: 33 4 67040231. E-mail: philips{at}igm.cnrs-mop.fr.

Molecular and Cellular Biology, November 2002, p. 7769-7779, Vol. 22, No. 22
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.22.7769-7779.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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