Isolation and characterization of the cDNAs encoding two isoforms of subunit CIX of bovine cytochrome c oxidase.
The smallest subunit of bovine cytochrome c oxidase (CIX or VIII in different nomenclatures) occurs in two isoforms, a heart (H) form and a liver (L) form. The cDNAs for both of these forms have been isolated and sequenced. The cDNA for the H form encodes a protein 70 amino acids long with a 24-residue presequence and a mature polypeptide of 46 amino acids; that of liver encodes a protein of 69 amino acids, a 25-residue presequence and a mature polypeptide of 44 amino acids. The leader sequences of the H and L forms are 40% homologous with an abundance of positively charged residues but no negatively charged amino acids. These features are typical of polypeptides targeted to the mitochondrion for processing in the matrix space. The homology of the two isoforms is 52% in the mature subunit with most of the differences occurring in the N-terminal hydrophilic domain of the protein. Evidence has been obtained of polymorphisms of both the H and L forms of the subunit. Protein chemical analyses show that the H isoform is the predominant if not the exclusive form of subunit CIX in heart and skeletal muscle tissue. The L form is the predominant form in liver, kidney, and brain. Northern analyses, using cDNAs to the two forms to screen whole cell RNA preparations, show that the transcript of the H isoform is present in heart and skeletal muscle but not in other tissues examined. The mRNA of the L form was found in brain, kidney, and liver and also in heart and skeletal muscle. These results indicate that the synthesis of the H isoform of CIX is controlled transcriptionally while the L form is under post-transcriptional regulation at least in heart and muscle tissue.
Lightowlers
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Institute of Molecular Biology, University of Oregon, Eugene 97403.
Ewart
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Aggeler
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Zhang
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Calavetta
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Capaldi
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002417:Cattle; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D003576:Electron Transport Complex IV / Q000235:genetics*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009928:Organ Specificity; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D009626:Terminology as Topic
D007527:Isoenzymes; D046911:Macromolecular Substances; D012313:RNA; D004247:DNA; D003576:Electron Transport Complex IV