A protein which binds preferentially to single-stranded core sequence of autonomously replicating sequence is essential for respiratory function in mitochondrial of Saccharomyces cerevisiae.
From yeast nuclear extract, we have identified several DNA-protein complexes using the T-rich strand of core consensus sequence of autonomously replicating sequence by gel shift assay. One of them showed preferential binding to the T-rich sequence of the DNA. We have partially purified a protein constituent of this complex and cloned its gene. The gene has an open reading frame encoding a protein of 380 amino acids (M(r) = 42,100) which is processed to a mature protein of 371 amino acids (M(r) = 40,900). The protein has neither significant amino acid homology with any previously reported proteins nor characteristic motifs. A putative HAP2/HAP3/HAP4 binding sequence was found at about 1 kilobase upstream of the gene. Disruption of the chromosomal gene revealed that the gene was neither essential for cell viability nor involved in DNA replication, but was essential for mitochondrial respiratory function. We therefore named the gene MRF1 for mitochondrial respiratory function 1. In a mrf1 null mutant the absorption spectra of cytochromes b, a, and a3 were undetectable, although mitochondrial DNA and protein synthesis in mitochondria were intact. Antibodies against MRF1 detected the antigen localized predominantly in the nucleus in vivo. These results suggest that MRF1 is a transcriptional regulatory protein of some genes whose products are necessary for the functional assembly of mitochondrial respiratory proteins.
Yamazoe
M
M
Department of Genetics, Osaka University Medical School, Japan.
Shirahige
K
K
Rashid
M B
MB
Kaneko
Y
Y
Nakayama
T
T
Ogasawara
N
N
Yoshikawa
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004261:DNA Replication; D004274:DNA, Recombinant / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000096:biosynthesis / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010100:Oxygen / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors
D004274:DNA, Recombinant; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C071720:MTF1 protein, S cerevisiae; D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D010100:Oxygen