Topogenesis of Cytochrome Oxidase Subunit II

Cytochrome c oxidase subunit II (COXII) in yeast mitochondria is synthesized as a precursor (preCOXII) and is sorted across the inner membrane, whereby both N and C termini become exposed to the intermembrane space. We describe here how this process can be experimentally dissected into a number of distinct stages. Our results demonstrate that the translation of COXII is not obligatorily coupled to translocation. Insertion into the inner membrane and export of the N- and C-terminal domains require an energized inner membrane. The export of COXII is independent of both maturation by the Imp1p protease and assembly into the cytochrome c oxidase complex. When linked to a mitochondrial matrix-targeting sequence, the N-terminal portion of preCOXII (fused to mouse dihydrofolate reductase) can be imported into the mitochondrial matrix. Following accumulation in the matrix, this chimeric protein can become exported across the inner membrane, delivering the N terminus into the intermembrane space where it undergoes processing by the Imp1p protease. This export process displays a number of similarities to bacterial protein export and supports the view that the principles of sorting are conserved from prokaryotes to eukaryotic organelles.

[1]  A. Gruhler,et al.  A novel intermediate on the import pathway of cytochrome b2 into mitochondria: evidence for conservative sorting. , 1995, The EMBO journal.

[2]  W. Neupert,et al.  Mitochondrial heat shock protein 70, a molecular chaperone for proteins encoded by mitochondrial DNA , 1994, The Journal of cell biology.

[3]  H. Feldmann,et al.  Yta10p is required for the ATP‐dependent degradation of polypeptides in the inner membrane of mitochondria , 1994, FEBS letters.

[4]  G. Heijne,et al.  Sec‐independent translocation of a 100‐residue periplasmic N‐terminal tail in the E. coli inner membrane protein proW. , 1994, The EMBO journal.

[5]  P. Slonimski,et al.  OXA1, a Saccharomyces cerevisiae nuclear gene whose sequence is conserved from prokaryotes to eukaryotes controls cytochrome oxidase biogenesis. , 1994, Journal of molecular biology.

[6]  W. Neupert,et al.  Targeting of cytochrome b2 into the mitochondrial intermembrane space: specific recognition of the sorting signal. , 1993, The EMBO journal.

[7]  R. O. Poyton,et al.  Protein export from the mitochondrial matrix. , 1992, Trends in cell biology.

[8]  P. Covello,et al.  Silent mitochondrial and active nuclear genes for subunit 2 of cytochrome c oxidase (cox2) in soybean: evidence for RNA‐mediated gene transfer. , 1992, The EMBO journal.

[9]  R. Hallberg,et al.  Cytochromes c 1 and b 2 are sorted to the intermembrane space of yeast mitochondria by a stop-transfer mechanism , 1992, Cell.

[10]  J. Palmer,et al.  RNA-mediated transfer of the gene coxII from the mitochondrion to the nucleus during flowering plant evolution , 1991, Cell.

[11]  P. Slonimski,et al.  ABC1, a novel yeast nuclear gene has a dual function in mitochondria: it suppresses a cytochrome b mRNA translation defect and is essential for the electron transfer in the bc 1 complex. , 1991, The EMBO journal.

[12]  Arnold J. M. Driessen,et al.  Δμ H+ and ATP function at different steps of the catalytic cycle of preprotein translocase , 1991, Cell.

[13]  Jon Beckwith,et al.  The role of charged amino acids in the localization of secreted and membrane proteins , 1990, Cell.

[14]  N. Capitanio,et al.  Cytochrome oxidase assembly in yeast requires the product of COX11, a homolog of the P. denitrificans protein encoded by ORF3. , 1990, The EMBO journal.

[15]  F. Nóbrega,et al.  COX10 codes for a protein homologous to the ORF1 product of Paracoccus denitrificans and is required for the synthesis of yeast cytochrome oxidase. , 1990, The Journal of biological chemistry.

[16]  R. Dalbey.  Positively charged residues are important determinants of membrane protein topology. , 1990, Trends in biochemical sciences.

[17]  S. Ackerman,et al.  ATP10, a yeast nuclear gene required for the assembly of the mitochondrial F1-F0 complex. , 1990, The Journal of biological chemistry.

[18]  M. Costanzo,et al.  Control of mitochondrial gene expression in Saccharomyces cerevisiae. , 1990, Annual review of genetics.

[19]  G. Vonheijne,et al.  Control of topology and mode of assembly of a polytopic membrane protein by positively charged residues , 1989, Nature.

[20]  F. Hartl,et al.  Protein folding in mitochondria requires complex formation with hsp60 and ATP hydrolysis , 1989, Nature.

[21]  R. O. Poyton,et al.  A role for membrane potential in the biogenesis of cytochrome c oxidase subunit II, a mitochondrial gene product. , 1989, The Journal of biological chemistry.

[22]  F. Hartl,et al.  Mitochondrial precursor proteins are imported through a hydrophilic membrane environment. , 1987, European journal of biochemistry.

[23]  W. Neupert,et al.  Import of cytochrome c into mitochondria , 1987 .

[24]  H. Fritz,et al.  Oligonucleotide-directed construction of mutations via gapped duplex DNA. , 1987, Methods in enzymology.

[25]  A. Myers,et al.  Genetics of mitochondrial biogenesis. , 1986, Annual review of biochemistry.

[26]  W. Dowhan,et al.  The cytoplasmically‐made subunit IV is necessary for assembly of cytochrome c oxidase in yeast. , 1985, The EMBO journal.

[27]  R. O. Poyton,et al.  Mitochondrial gene expression in saccharomyces cerevisiae. I. Optimal conditions for protein synthesis in isolated mitochondria. , 1984, The Journal of biological chemistry.

[28]  G. Macino,et al.  Cytochrome oxidase subunit 2 gene in Neurospora crassa mitochondria. , 1983, The Journal of biological chemistry.

[29]  G. Mannhaupt,et al.  A nuclear mutation prevents processing of a mitochondrially encoded membrane protein in Saccharomyces cerevisiae. , 1983, The EMBO journal.

[30]  R. Bisson,et al.  Localization of lipid binding domain(s) on subunit II of beef heart cytochrome c oxidase. , 1982, The Journal of biological chemistry.

[31]  H. Shio,et al.  Polypeptide and phospholipid composition of the membrane of rat liver peroxisomes: comparison with endoplasmic reticulum and mitochondrial membranes , 1982, The Journal of cell biology.

[32]  M. Hunkapiller,et al.  Alignment of the amino terminal amino acid sequence of human cytochrome c oxidase subunits I and II with the sequence of their putative mRNAs. , 1981, Nucleic acids research.

[33]  R. O. Poyton,et al.  Mitochondrial membrane biogenesis: identification of a precursor to yeast cytochrome c oxidase subunit II, an integral polypeptide. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[34]  H. Nau,et al.  Studies on cytochrome c oxidase, VII. Isolation and chemical characterization of polypeptide VII. , 1979, Hoppe-Seyler's Zeitschrift fur physiologische Chemie.

[35]  L. Grivell,et al.  The mitochondrial genome of yeast , 1978, Cell.

[36]  H. Pelham,et al.  An efficient mRNA-dependent translation system from reticulocyte lysates. , 1976, European journal of biochemistry.

[37]  G. Schatz,et al.  Cytochrome c oxidase from bakers' yeast. II. Site of translation of the protein components. , 1973, The Journal of biological chemistry.

[38]  K. Watson.  THE ORGANIZATION OF RIBOSOMAL GRANULES WITHIN MITOCHONDRIAL STRUCTURES OF AEROBIC AND ANAEROBIC CELLS OF SACCHAROMYCES CEREVISAE , 1972, The Journal of cell biology.

[39]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[40]  J. Huet,et al.  Isolation and characterization of ribosomes from yeast mitochondria , 1969, FEBS letters.