OXBOX, a positive transcriptional element of the heart-skeletal muscle ADP/ATP translocator gene.
Three positive transcriptional control regions have been identified in the promoter of the human heart-skeletal muscle adenine nucleotide translocator gene (ANT1). By transfecting promoter-chloramphenicol acetyltransferase fusion constructs into C2C12 myogenic cells, each positive region was found to increase transcription 2-3-fold. The first region spans from -123 to -674 base pairs (bp), the second from -2.6 to -3.1 kilobases, and the third from -3.1 to -8.8 kilobases. Linker-scanning mutants generated using the polymerase chain reaction and modified oligonucleotides have identified the OXBOX (5'-GGCTCTAAAGAGG) as the positive element within the -123 to -674-bp region. This element enhances transcription in muscle cells but not in HeLa cells, suggesting that it is muscle-specific. Gel retardation experiments have revealed a factor from C2C12 cells which specifically binds to a 40-bp piece of the ANT1 promoter containing the OXBOX. Since the OXBOX is also found in the promoter of the human ATP synthase beta subunit gene, it is the first tissue-specific element identified which could coordinately regulate mitochondrial oxidative phosphorylation genes.
Li
K
K
Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia 30322.
Hodge
J A
JA
Wallace
D C
DC
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002872:Chromosome Deletion; D005796:Genes; D051379:Mice; D000226:Mitochondrial ADP, ATP Translocases / Q000235:genetics*; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology*; D016297:Mutagenesis, Site-Directed; D009206:Myocardium / Q000201:enzymology*; D015345:Oligonucleotide Probes; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014162:Transfection
D015345:Oligonucleotide Probes; D000226:Mitochondrial ADP, ATP Translocases; D015500:Chloramphenicol O-Acetyltransferase