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(Received for publication, March 7, 1995; and in revised form, May 26, 1995) From the Nuclear respiratory factor 1 (NRF-1) is a transcription factor
that acts on nuclear genes encoding respiratory subunits and components
of the mitochondrial transcription and replication machinery. Here we
describe the isolation and characterization of the human gene encoding
NRF-1. The human genomic sequences detected with NRF-1 cDNA probes at
high stringency are all contained within seven overlapping recombinant
Volume 270,
Number 30,
Issue of July 30, pp. 18019-18025, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
clones. The NRF-1 gene encompassed by these recombinants spans
65 kilobases (kb) and has 11 exons and 10 introns that range in
size from 0.8 to 15 kb. A rapid amplification of cDNA ends-polymerase
chain reaction product containing the 5`-terminus of the NRF-1 cDNA has
two exons from the 5`-untranslated region and terminates at a major
transcription initiation site identified by S1 nuclease mapping. A
genomic fragment containing a portion of the 5`-terminal exon and an
additional 1 kb upstream had a functional promoter that was active in
transfected COS cells, HeLa cells, and L6 myoblasts. The transcription
initiation site utilized by the transfected promoter corresponded to
that used by the endogenous gene in vivo. NRF-1 mRNA was
expressed at very low levels in rat tissues compared with cytochrome c and, unlike cytochrome c, was most abundantly
expressed in lung and testis. The NRF-1 gene was localized to human
chromosome 7 by analysis of DNA from a panel of human-hamster cell
hybrids with human-specific NRF-1 polymerase chain reaction primers.
This assignment was further refined to 7q31 by cohybridization of
NRF-1- and chromosome 7-specific probes to human metaphase chromosomes.
These analyses should be useful in evaluating the potential role of
NRF-1 in mitochondrial diseases resulting from defects in the nuclear
control of mitochondrial function.
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