Characterization and identification of promoter elements in the mouse COX17 gene.
Cox17p, essential for the assembly of functional cytochrome c oxidase (CCO) in Saccharomyces cerevisiae, has been believed to deliver copper ions to the mitochondrion for insertion into the enzyme. We have recently isolated an approximately 20 kb genomic fragment of the mouse COX17. Reporter assay experiments have shown that most of the promoter activity was restricted to a 0.85 kb fragment flanking the first exon. Further intensive deletion and detailed mutation analysis suggested that the minimal essential region for transactivation was located at bases -155 to -70. This 5'-flanking region did not possess a TATA box, but contained putative Sp1, NRF-1 and NRF-2 binding sites. COX17 basal promoter activity was abrogated by site-directed mutagenesis of Sp1, NRF-1 and NRF-2 binding sites. Electrophoretic mobility shift assays with AtT-20 and NIH3T3 cell nuclear extract revealed that this region binds both a Sp1-like protein and NRF-1 transcription factors. These results indicated that Sp1, NRF-1 and NRF-2 are involved in basal transcription of the COX17 gene, similar to the transcription mechanism of other CCO-related genes.
Takahashi
Yoshinori
Y
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki 305-8572, Tsukuba, Japan.
Kako
Koichiro
K
Arai
Hidenori
H
Ohishi
Takahiro
T
Inada
Yoshiko
Y
Takehara
Akio
A
Fukamizu
Akiyoshi
A
Munekata
Eisuke
E
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D027682:Cation Transport Proteins / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D000079922:Copper Transport Proteins; D004268:DNA-Binding Proteins / Q000378:metabolism; D051268:GA-Binding Protein Transcription Factor; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D051265:NF-E2-Related Factor 1; D051266:Nuclear Respiratory Factor 1; D051264:Nuclear Respiratory Factors; D011401:Promoter Regions, Genetic; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D024363:Transcription Initiation Site; D014162:Transfection
D027682:Cation Transport Proteins; D000079922:Copper Transport Proteins; C491005:Cox17 protein, mouse; D004268:DNA-Binding Proteins; D051268:GA-Binding Protein Transcription Factor; D008565:Membrane Proteins; D051265:NF-E2-Related Factor 1; C495634:Nrf1 protein, mouse; D051266:Nuclear Respiratory Factor 1; D051264:Nuclear Respiratory Factors; D015534:Trans-Activators; D014157:Transcription Factors