CDC6 expression is regulated by lineage-specific transcription factor GATA1

• Published in: Cell cycle (Georgetown, Tex.) Tex.), 2012-08, Vol.11 (16), p.3055-3066
• Main Authors: Fernández-Morales, Bárbara, Pavón, Leticia, Calés, Carmela
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Binding; Binding Sites; Biology; Bioscience; Calcium; Cancer; CDC6; Cell; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cell Differentiation; Cell Lineage; Cell Proliferation; Chromatin Immunoprecipitation; Conserved Sequence; CP2; Cycle; Electrophoretic Mobility Shift Assay; Enhancer Elements, Genetic; GATA1; GATA1 Transcription Factor - genetics; GATA1 Transcription Factor - metabolism; GATA1 Transcription Factor - pharmacology; GATA1s; Gene Expression Regulation; Genes, Reporter; HEK293 Cells; Humans; Landes; megakaryocytes; Megakaryocytes - cytology; Megakaryocytes - metabolism; Mice; Molecular Sequence Data; NIH 3T3 Cells; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Organogenesis; Proteins; Retroviridae - genetics; Retroviridae - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Transcription Initiation Site; Transcription, Genetic; Transcriptional Activation; transcriptional regulation; Transfection
• ISSN: 1538-4101; 1551-4005
• DOI: 10.4161/cc.21471

GATA1 is a hematopoietic transcription factor essential for expression of most genes encoding erythro-megakaryocytic proteins, i.e., globins and platelet glycoproteins. A role for GATA1 as a cell proliferation regulator has been proposed, as some of its bona fide targets comprise global regulators, such as c-KIT or c-MYC, or cell cycle factors, i.e., CYCLIN D or p21CIP1. In this study, we describe that GATA1 directly regulates the expression of replication licensing factor CDC6. Using reporter transactivation, electrophoretic mobility shift and chromatin immunoprecipitation assays, we show that GATA1 stimulates CDC6 transcription by binding to a canonical binding site located within a 166bp enhancer region upstream CDC6 promoter. This evolutionary conserved GATA binding site conforms to recently described chromatin occupancy rules, i.e., preferred bases within core WGATAR (TGATAA), 5' and 3' flanking bases (GGTGATAAGG) and distance to the transcription initiation site. We also found adjacent conserved binding sites for ubiquitously expressed transcription factor CP2, needed for GATA activity on CDC6 enhancer. Our results add to the growing evidence for GATA1 acting as a direct transcriptional regulator of the cell cycle machinery, thus linking cell proliferation control and specific gene expression programs during lineage differentiation.