The Dicyclohexylcarbodiimide-Binding Protein of Rat Liver Mitochondria as a Product of the Mitochondrial Protein Synthesis

Dianoux, Anne-Christine; Bof, Mireille; Vignais, Pierre V.

doi:10.1111/j.1432-1033.1978.tb12423.x

Cited by 25 publications

(3 citation statements)

References 33 publications

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“…The biogenesis of this subunit has been studied in yeast (2, 5), Neurospora (1), and in higher eukaryotes (15,16). In yeast this polypeptide is coded for by a mitochondrial gene, which has been isolated and sequenced, and is translated on mitochondrial ribosomes (17,18).…”

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Biosynthetic pathway of mitochondrial ATPase subunit 9 in Neurospora crassa.

Schmidt¹,

Hennig²,

Zimmermann³

et al. 1983

The Journal of Cell Biology

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Subunit 9 of mitochondriat ATPase (Su9) is synthesized in reticulocyte lysates programmed with Neurospora poly A-RNA, and in a Neurospora cell free system as a precursor with a higher apparent molecular weight than the mature protein (Mr 16,400 vs. 10,500). The RNA which directs the synthesis of Su9 precursor is associated with free polysomes. The precursor occurs as a high molecular weight aggregate in the postribosomal supernatant of reticulocyte lysates. Transfer in vitro of the precursor into isolated mitochondria is demonstrated. This process includes the correct proteolytic cleavage of the precursor to the mature form. After transfer, the protein acquires the following properties of the assembled subunit: it is resistant to added protease, it is soluble in chloroform/methanol, and it can be immunoprecipitated with antibodies to FI-ATPase. The precursor to Su9 is also detected in intact cells after pulse labeling. Processing in vivo takes place posttranslationally. It is inhibited by the uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP).A hypothetical mechanism is discussed for the intracellular transfer of Su9. It entails synthesis on free polysomes, release of the precursor into the cytosol, recognition by a receptor on the mitochondrial surface, and transfer into the inner mitochondrial membrane, which is accompanied by proteolytic cleavage and which depends on an electrical potential across the inner mitochondrial membrane.

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confidence: 99%

Biosynthetic pathway of mitochondrial ATPase subunit 9 in Neurospora crassa.

Schmidt¹,

Hennig²,

Zimmermann³

et al. 1983

The Journal of Cell Biology

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“…Further confirmation of the site of synthesis of this protein is given by the fact that the N terminus is not formylmethionine, as is the case for the dicyclohexylcarbodiimide-binding component of yeast [14]. Recently, it has been reported that the dicyclohexylcarbodiimide-binding protein of rat liver mitochondria is a mitochondria1 product, though the protein was not clearly resolved on gels, and end group determination was not carried out as a check for purity [32]. It is possible that a low-molecular-weight mitochondrial translation product(s), extractable from a trichloroacetic acid precipitate of whole mitochondria by chloroform/methanol and similar to that of A .…”

Section: Dicyclohexylcarbodiimide-binding Proteinmentioning

confidence: 99%

Mitochondrial ATPase Complex of Aspergillus nidulans and the Dicyclohexylcarbodiimide‐Binding Protein

Turner

Imam

Küntzel

1979

European Journal of Biochemistry

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The dicyclohexylcarbodiimide-binding protein of Aspergillus nidulans has been identified as the smallest subunit of the mitochondrial ATPase complex, and has a molecular weight of approximately 8000. It is extractable from whole mitochondria and from the purified enzyme in neutral chloroform/ methanol, contains 30% polar amino acids, and the N-terminal amino acid has been identified as tyrosine. Using a double-labelling technique in the absence and presence of cycloheximide, followed by immunoprecipitation of the enzyme complex with antiserum against Neurospora crassa F1 ATPase, it has been shown that this subunit is synthesized on cytoplasmic ribosomes.Oligomycin-resistant mutants have been isolated in the filamentous Ascomycete Aspergillus nidulans [l ] and mutations in both nuclear and extranuclear genomes have been shown to affect the oligomycin sensitivity of the mitochondria1 ATPase activity [2,3], suggesting the involvement of at least two subunits of the enzyme complex, one nuclearly coded and one extranuclearly coded, in the expression of oligomycin resistance. Furthermore, nuclear-extranuclear interactions have been observed in strains containing mutations in both genomes [3]. In contrast, only extranuclear oligomycin-resistant mutants in Saccharomyces cerevisiae [4,5] and nuclear oligomycin-resistant mutants in Neurospora crassa [6] have been shown to alter the resistance of the ATPase activity of these organisms in vitro.In order to identify the lesions caused by these mutants, and thus the gene-protein relationships, studies on the ATPase complex are essential. Oligomycin is known to inhibit oxidative phosphorylation by acting on the Fo or membrane components of the complex [7]. Dicyclohexylcarbodiimide acts at a nearby site in a similar manner, and shows irreversible binding to one of the FO components [8-111. This hydrophobic polypeptide, referred to as the dicyclohexylcarbodiimide-binding protein, has now been extracted from a number of sources, including the mitochondria of beef heart [ll, 121, S. cerevisiae [13,14] and N . crassa 11.51, the chloroplasts of lettuce [16] and from membranes of Escherichia coli [17]. In all of these species the protein has a molecular weight of

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“…These studies confirm earlier work on the biogenesis of the ATPase in these THE JOURNAL OF CELL BIOLOGY " VOLUME 86 SEPTEMBER 1980 723-729 © The Rockefeller University Press -0021-9525/80/09/0723/07 $1 .00 organisms (23,49,50) . There are conflicting reports on the site of biosynthesis of the DCCD-binding protein in animal cells (1, 12,13,25).…”

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confidence: 99%