Nuclear respiratory factor 1 activation sites in genes encoding the gamma-subunit of ATP synthase, eukaryotic initiation factor 2 alpha, and tyrosine aminotransferase. Specific interaction of purified NRF-1 with multiple target genes.
Transcription factor nuclear respiratory factor 1 (NRF-1) was originally identified as an activator of the cytochrome c gene and subsequently found to stimulate transcription through specific sites in other nuclear genes whose products function in the mitochondria. These include subunits of the cytochrome oxidase and reductase complexes and a component of the mitochondrial DNA replication machinery. Here we establish that a functional recognition site for NRF-1 is present in the ATP synthase gamma-subunit gene extending the proposed respiratory role of NRF-1 to complex V. In addition, biologically active NRF-1 sites are found in genes encoding the eukaryotic translation initiation factor 2 alpha-subunit and tyrosine aminotransferase, both of which participate in the rate-limiting step of their respective pathways of protein biosynthesis and tyrosine catabolism. The recognition sites from each of these genes form identical complexes with NRF-1 as established by competition binding assays, methylation interference footprinting, and UV-induced DNA cross-linking. Cloned oligomers of each NRF-1 binding site also stimulate the activity of a truncated cytochrome c promoter in transfected cells. The NRF-1 binding activities for the various target sites copurified approximately 33,000-fold and resided in a single protein of 68 kDa. These observations further support a role for NRF-1 in the expression of nuclear respiratory genes and suggest it may help coordinate respiratory metabolism with other biosynthetic and degradative pathways.
Department of Cell, Molecular, and Structural Biology, Northwestern University Medical School, Chicago, Illinois 60611.
C M
CM
Chau
M J
MJ
Evans
R C
RC
Scarpulla
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D004272:DNA, Mitochondrial / Q000235:genetics / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008745:Methylation; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D006180:Proton-Translocating ATPases / Q000235:genetics*; D051381:Rats; D014157:Transcription Factors / Q000378:metabolism*; D014162:Transfection; D014444:Tyrosine Transaminase / Q000235:genetics*; D014466:Ultraviolet Rays
D004272:DNA, Mitochondrial; D004268:DNA-Binding Proteins; D015852:Eukaryotic Initiation Factor-2; D014157:Transcription Factors; D014444:Tyrosine Transaminase; D006180:Proton-Translocating ATPases