Molecular cloning of a cDNA for the smallest nuclear-encoded subunit of sweet potato cytochrome c oxidase. Analysis with the cDNA of the structure and import into mitochondria of the subunit.
A cDNA clone for the subunit Vc, the smallest nuclear-encoded subunit, of sweet potato cytochrome c oxidase was isolated. Nucleotide sequence analysis showed that the subunit is synthesized as a precursor from which only the amino-terminal amino acid, methionine, residue is removed to form the mature subunit. The mature subunit consists of 63 amino acid residues which are arranged so as to form one hydrophobic cluster lying between two hydrophilic clusters; such an arrangement is also seen with some nuclear-encoded small subunits of the enzymes from other eukaryotes. The labelled precursor protein was synthesized with the cDNA by in vitro transcription (SP6 system) and translation (wheat germ system). This precursor was imported into isolated sweet potato tuberous root mitochondria without any significant processing, the import being dependent on membrane potential.
Nakagawa
T
T
Laboratory of Biochemistry, School of Agriculture, Nagoya University, Japan.
Maeshima
M
M
Nakamura
K
K
Asahi
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics* / Q000378:metabolism; D019286:Endopeptidase K; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010944:Plants / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases / Q000378:metabolism; D014158:Transcription, Genetic; D014675:Vegetables / Q000201:enzymology / Q000235:genetics
D015345:Oligonucleotide Probes; D011498:Protein Precursors; D012333:RNA, Messenger; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D004247:DNA; D003576:Electron Transport Complex IV; D012697:Serine Endopeptidases; D019286:Endopeptidase K