The NRF-1/alpha-PAL transcription factor regulates human E2F6 promoter activity.
E2F6 is widely expressed in human tissues and cell lines. Recent studies have demonstrated its involvement in developmental patterning and in the regulation of various genes implicated in chromatin remodelling. Despite a growing number of studies, nothing is really known concerning the E2F6 expression regulation. To understand how cells control E2F6 expression, we analysed the activity of the previously cloned promoter region of the human E2F6 gene. DNase I footprinting, gel electrophoreticmobility shift, transient transfection and site-directed mutagenesis experiments allowed the identification of two functional NRF-1/alpha-PAL (nuclear respiratory factor-1/alpha-palindrome-binding protein)-binding sites within the human E2F6 core promoter region, which are conserved in the mouse and rat E2F6 promoter region. Moreover, ChIP (chromatin immunoprecipitation) analysis demonstrated that overexpressed NRF-1/alpha-PAL is associated in vivo with the E2F6 promoter. Furthermore, overexpression of full-length NRF-1/alpha-PAL enhanced E2F6 promoter activity, whereas expression of its dominant-negative form reduced the promoter activity. Our results indicate that NRF-1/alpha-PAL is implicated in the regulation of basal E2F6 gene expression.
Kherrouche
Zoulika
Z
CNRS UMR 8117, Institut de Biologie de Lille, 1 rue Calmette, BP 447, 59021 Lille, France.
De Launoit
Yvan
Y
Monte
Didier
D
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics / Q000502:physiology; D001859:Bone Neoplasms / Q000473:pathology; D001943:Breast Neoplasms / Q000473:pathology; D045744:Cell Line, Tumor; D017124:Conserved Sequence / Q000235:genetics / Q000502:physiology; D018983:DNA Footprinting / Q000379:methods; D004273:DNA, Neoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000502:physiology*; D003850:Deoxyribonuclease I / Q000378:metabolism; D050701:E2F6 Transcription Factor; D005260:Female; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D051265:NF-E2-Related Factor 1; D051266:Nuclear Respiratory Factor 1; D051264:Nuclear Respiratory Factors; D012516:Osteosarcoma / Q000473:pathology; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011485:Protein Binding / Q000235:genetics / Q000502:physiology; D051381:Rats; D016415:Sequence Alignment / Q000379:methods; D015534:Trans-Activators / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics* / Q000502:physiology*; D015533:Transcriptional Activation / Q000235:genetics
D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D050701:E2F6 Transcription Factor; C491744:E2F6 protein, human; D051265:NF-E2-Related Factor 1; C495633:NRF1 protein, human; C495632:Nfe2l1 protein, rat; C495634:Nrf1 protein, mouse; D051266:Nuclear Respiratory Factor 1; D051264:Nuclear Respiratory Factors; D015534:Trans-Activators; D014157:Transcription Factors; C088388:alpha-Pal transcription factor; D003850:Deoxyribonuclease I