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Originally published In Press as doi:10.1074/jbc.M909390199 on March 30, 2000

J. Biol. Chem., Vol. 275, Issue 25, 19258-19267, June 23, 2000
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DcE2F, a Functional Plant E2F-like Transcriptional Activator from Daucus carota*

Diego AlbaniDagger §, Luisa Mariconti, Stefano Ricagno, Letizia Pitto||, Cristina Moroni**, Kristian Helin**, and Rino Cella

From the Dagger  Department of Botany and Plant Ecology, University of Sassari, Via Muroni 25, 07100 Sassari, the  Department of Genetics and Microbiology, University of Pavia, Via Ferrata 1, 27100 Pavia, the || Istituto di Mutagenesi e Differenziamento, Via Svezia 10, 56124 Pisa, and the ** European Institute of Oncology, Department of Experimental Oncology, Via Ripamonti 435, 20141 Milan, Italy

In animal cells the progression of the cell cycle through G1/S transition and S phase is under the control of the pRB/E2F regulatory pathway. The E2F transcription factors are key activators of genes coding for several regulatory proteins and for enzymes involved in nucleotide and DNA synthesis. In this report we have detected the presence of E2F-like DNA binding activities in carrot nuclear extracts, and we have isolated a carrot cDNA (DcE2F) encoding a plant E2F homologue. The DcE2F gene is expressed in proliferating cells and is induced during the G1/S transition of the cell cycle. Supershift experiments using anti-DcE2F antiserum have confirmed that the DcE2F protein is a component of the carrot E2F-like nuclear activities. DNA binding assays have demonstrated that the DcE2F protein can recognize a canonical E2F cis-element in association with a mammalian DP protein. Furthermore, transactivation assays have revealed that DcE2F is a functional transcription factor that can transactivate, together with a DP partner, an E2F-responsive reporter gene in both plant and mammalian cells.

* This work was supported by grants of the University of Pavia "Progetto di Ricerca di Ateneo" and by "Programmi di Ricerca Scientifica di Interesse Nazionale" of Ministero dell' Università e della Ricerca Scientifica e Tecnologica.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AJ251586.

§ To whom correspondence should be addressed. Tel.: 39-79-228645; Fax: 39-79-233600; E-mail: albani@ssmain.uniss.it.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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