Cell-Free Synthesis of a Larger-Molecular-Weight Precursor of Cytochrome c Oxidase Subunit V from Rat Liver and the Distribution of Its mRNA between Free and Membrane-Bound Polysomes

Schmelzer, Elmon; Northemann, Wolfgang; Kadenbach, Bernhard; Heinrich, Peter C.

doi:10.1111/j.1432-1033.1982.tb06853.x

Cited by 12 publications

(1 citation statement)

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“…However, since the reading frame in pCOX4.419 remains open upstream for ten additional' codons to thr end of the cDNA insert, and none of the ten encodes methionine, we cannot rule out a larger extension. Estimates of the presequence length pf another cytochrome oxidase component, subunit V,, in yeast (11)' and rat (6) provide similar values of about 20-25 residues.…”

Section: Resultsmentioning

confidence: 84%

Isolation and characterization of a cDNA clone for bovine cytochrome c oxidase subunit IV.

Lomax¹,

Bachman

Nasoff

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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We have isolated a cDNA clone for the precursor to subunit IV of bovine cytochrome c oxidase (ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1). A eDNA library was constructed from poly(A)+ RNA of adult beef liver by insertion of cDNA into the plasmid vector pBR322. Transformants were screened by colony hybridization with two mixtures of [32P]-labeled synthetic oligodeoxyribonucleotioces. We screened 20,000 transformants with a mixture of heptadecamers complementary to all 16 possible sequences encoding amino acids 98-103 and obtained two cDNA clones encoding subunit IV amino acid sequences. We determined the DNA sequence of the larger (416 base-pair) insert, which contains the coding sequence for amino acids 1-107 of the mature protein and an NH2-terminal extension (presequence). The deduced amino acid sequence of the mature protein is identical with the previously determined protein sequence; the sequence of the N112-terminal extension contains a potential initiator methionine at amino acid -22 from the NH2-terminus of the processed protein. The presequence is quite basic and contains several arginines, including one at the processing site. No hydrophobic region analogous to that found in bacterial and eukaryotic signal peptides is present, but there are homologies with other mitochondrial protein presequences, which may include a common signal for their destination and processing.Cytochrome c oxidase (cytochrome oxidase; ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1), a mnultisubunit enzyme complex located in the mitochondrial inner membrane, transfers electrons to oxygen in the terminal reaction of the electron transport chain. The enzyme isolated from beef heart is composed of at least seven subunits (reviewed in ref. 1). Like several other mitochondrial complexes, cytochrome oxidase is the product of two separate genomes. The three largest subunits (1-111) are encoded by mitochondrial DNA (2) and synthesized on mitochondrial ribosomes (3, 4), whereas the smaller subunits (IV-VII) are nuclear gene products. Subunits IV, V, and VI are synthesized on cytoplasmic ribosomes as precursor molecules containing NH2-terminal extensions (presequernces) (5-9), transported to mitochondria, and processed into mature subunits by 'a mitochondrial protease (reviewed in ref. 10; see also refs. 11-13). Little is known of either the signals that direct these proteins to mitochondria or the structural requirements for their correct processing.To investigate both nuclear-cytoplasmic interactions and the rate of evolution of nuclear genes for mitochondrial proteins, we are cloning the nuclear genes for cytochrome c oxidase. Since the amino acid sequences of most of the nuclearcoded bovine cytochrome oxidase subunits are known (1), it 6295The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Section: Resultsmentioning

confidence: 84%

Isolation and characterization of a cDNA clone for bovine cytochrome c oxidase subunit IV.

Lomax¹,

Bachman

Nasoff

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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Struktur und Evolution des Atmungsferments Cytochrom‐c‐Oxidase

Kadenbach¹

1983

Angewandte Chemie

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Zum Andenken an Otto Warburgs ,100. GeburtstagVor ca. 50 Jahren erhielt Otto Warburg fur seine grundlegenden Arbeiten iiber das ,,Atmungsferment" den Nobelpreis. Doch erst gegen Ende der funfziger Jahre gelang die Isolierung dieses komplizierten Membranenzyms, das fur Atmung und Energiegewinnung der meisten Lebewesen auf der Erde unentbehrlich ist. Seitdem wurden intensive Untersuchungen durchgefuhrt, um den Mechanismus der Sauerstoffreduktion zu Wasser und die daran gekoppelte Protonentranslokation uber die Membran, die zur ATP-Synthese dient, zu verstehen. Die Erkenntnisse sind bisher allerdings unbefriedigend, nicht zuletzt deshalb, weil die vier Schwermetall-Redoxzentren an Proteine gebunden sind, deren Struktur zu erforschen erst in jiingster Zeit begonnen wurde. Dabei entdeckte man, daB Cytochrom-c-Oxidase aus Bakterien nur zwei oder drei, der tierische Enzymkomplex dagegen dreizehn Protein-Untereinheiten enthiilt. In diesem Beitrag werden die moglichen Funktionen der einzelnen Protein-Untereinheiten erlirtert ; am Beispiel der Cytochrom-c-Oxidase wird gezeigt, da13 die biochemische Evolution eine Zunahme an regulatorischer Kapazitat mit sich bringt.Angew. Chem. 95 (1983) 273-281 Q Verlag Chemie GmbH. 0-6940 Weinheim, 1983 00444249/83/0404-0273 S 02.50/0 213 Angew. G e m . 95 (1983) 281-313 Q Verlag Chemie GmbH. 0-6940 Weinheim, 1983 0044-8249/83/0404-028I S 02.50/0

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Structure and Evolution of the “Atmungsferment” Cytochrome c Oxidase

Kadenbach

1983

Angew. Chem. Int. Ed. Engl.

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Approximately 50 years ago, Otto Warburg received the Nobel Prize for his fundamental work on the “Atmungsferment”. But not until the end of the fifties was it possible to isolate this complicated membrane enzyme, which is necessary for respiration and energy production in most living organisms on earth. Since then, intensive research has been performed to elucidate the mechanism of reduction of oxygen to water and the coupled translocation of protons across the membrane which is involved in ATP synthesis. Until now the results have been unsatisfactory because the four catalytic heavy‐metal redox centers are bound to proteins, the structures of which have begun to be studied only recently. In the course of this research, it was discovered that cytochrome c oxidase from bacteria contains only two or three, whereas the enzyme complex from animals contains thirteen different protein components. The present article analyzes the possible functions of the various protein subunits and, using the example of cytochrome c oxidase, shows that biochemical evolution proceeds in such a way as to increase regulatory capacity.

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Cell-Free Synthesis of a Larger-Molecular-Weight Precursor of Cytochrome c Oxidase Subunit V from Rat Liver and the Distribution of Its mRNA between Free and Membrane-Bound Polysomes

Cited by 12 publications

References 42 publications

Isolation and characterization of a cDNA clone for bovine cytochrome c oxidase subunit IV.

Isolation and characterization of a cDNA clone for bovine cytochrome c oxidase subunit IV.

Struktur und Evolution des Atmungsferments Cytochrom‐c‐Oxidase

Structure and Evolution of the “Atmungsferment” Cytochrome c Oxidase

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