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A novel E2 box-GATA element modulates Cdc6 transcription during human cells polyploidization

AuthorsVilaboa, Nuria; Bermejo, Rodrigo ; Martinez, Pilar; Bornstein, Belén ; Calés, Carmela
Issue Date2004
PublisherOxford University Press
CitationNucleic Acids Research 32(21): 6454-6467 (2004)
AbstractCdc6 is a key regulator of the strict alternation of S and M phases during the mitotic cell cycle. In mammalian and plant cells that physiologically become polyploid, cdc6 is transcriptionally and post-translationally regulated. We have recently reported that Cdc6 levels are maintained in megakaryoblastic HEL cells, but severely downregulated by ectopic expression of transcriptional repressor Drosophila melanogaster escargot. Here, we show that cdc6 promoter activity is upregulated during megakaryocytic differentiation of HEL endoreplicating cells, and that Escargot interferes with such activation. Transactivation experiments showed that a 1.7 kb region located at 2800 upstream cdc6 transcription initiation site behaved as a potent enhancer in endoreplicating cells only. This activity was mainly dependent on a novel cis-regulatory element composed by an E2 box overlapping a GATA motif. Ectopic Escargot could bind this regulatory element in vitro and endogenous GATA-1 and E2A formed specific complexes in megakaryoblastic cells as well as in primary megakaryocytes. Chromatin Immunoprecipitation analysis revealed that both transcription factors were occupying the E2 box/GATA site in vivo. Altogether, these data suggest that cdc6 expression could be actively maintained during megakaryocytic differentiation through transcriptional mechanisms involving specific cis- and trans-regulatory elements. © Oxford University Press 2004; all rights reserved.
Publisher version (URL)http://dx.doi.org/10.1093/nar/gkh981
Identifiersdoi: 10.1093/nar/gkh981
issn: 0305-1048
e-issn: 1362-4962
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