Distinct functional contributions of 2 GABP-NRF-2 recognition sites within the context of the human TOMM70 promoter.
TOMM70 is a subunit of the outer mitochondrial membrane translocase that plays a major role as a receptor of hydrophobic preproteins targeted to mitochondria. We have previously reported 2 binding sites for the transcription factor GABP-NRF-2 in the promoter region of the human TOMM70 gene that are important in activating transcription. To assess the functionality and actual role of these sites, chromatin immunoprecipitation, site-directed mutagenesis, and electrophoretic mobility shift assays were carried out. We conclude that GABP-NRF-2 binds in vivo to the TOMM70 promoter, and that the 2 GABP-NRF-2 binding sites of the promoter have different functional contributions in promoting TOMM70 expression. Evidence is provided that they work in an additive manner as single sites.
Blesa
José R
JR
Fundación Centro de Investigación Príncipe Felipe, Av. Autopista del Saler 16-3, 46013 Valencia, Spain.
Hernández-Yago
José
J
D001483:Base Sequence; D001665:Binding Sites; D017124:Conserved Sequence; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000378:metabolism; D051268:GA-Binding Protein Transcription Factor / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics*; D033681:Mitochondrial Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D017354:Point Mutation; D011401:Promoter Regions, Genetic; D050784:Proto-Oncogene Protein c-ets-1 / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D051268:GA-Binding Protein Transcription Factor; D026901:Membrane Transport Proteins; D033681:Mitochondrial Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins; D009687:Nuclear Proteins; D050784:Proto-Oncogene Protein c-ets-1; C402118:TOMM70 protein, human