title: The 30-kilodalton subunit of bovine mitochondrial complex I is homologous to a protein coded in chloroplast DNA.
abstract: In cattle, 7 of the 30 or more subunits of the respiratory enzyme NADH:ubiquinone reductase (complex I) are encoded in mitochondrial DNA, and potential genes (open reading frames, orfs) for related proteins are found in the chloroplast genomes of Marchantia polymorpha and Nicotiana tabacum. Homologues of the nuclear-coded 49- and 23-kDa subunits are also coded in chloroplast DNA, and these orfs are clustered with four of the homologues of the mammalian mitochondrial genes. These findings have been taken to indicate that chloroplasts contain a relative of complex I. The present work provides further support. The 30-kDa subunit of the bovine enzyme is a component of the iron-sulfur protein fraction. Partial protein sequences have been determined, and synthetic oligonucleotide mixtures based on them have been employed as hybridization probes to identify cognate cDNA clones from a bovine library. Their sequences encode the mitochondrial import precursor of the 30-kDa subunit. The mature protein of 228 amino acids contains a segment of 57 amino acids which is closely related to parts of proteins encoded in orfs 169 and 158 in the chloroplast genomes of M. polymorpha and N. tabacum. Moreover, the chloroplast orfs are found near homologues of the mammalian mitochondrial genes for subunit ND3. Therefore, the plant chloroplast genomes have at least two separate clusters of potential genes encoding homologues of subunits of mitochondrial complex I. The bovine 30-kDa subunit has no extensive sequences of hydrophobic amino acids that could be folded into membrane-spanning alpha-helices, and although it contains two cysteine residues, there is no clear evidence in the sequence that it is an iron-sulfur protein.
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge, U.K.
authors.forename: S J
authors.initials: SJ
authors.lastname: Pilkington
authors.forename: J M
authors.initials: JM
authors.lastname: Skehel
authors.forename: J E
authors.initials: JE
authors.lastname: Walker
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002736:Chloroplasts / Q000378:metabolism*; D003001:Cloning, Molecular; D004272:DNA, Mitochondrial / Q000235:genetics*; D015723:Gene Library; D017343:Genes, Plant; D046911:Macromolecular Substances; D008929:Mitochondria, Heart / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016660:NAD(P)H Dehydrogenase (Quinone); D016366:Open Reading Frames; D010944:Plants / Q000235:genetics; D011808:Quinone Reductases / Q000235:genetics* / Q000302:isolation & purification; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
chemical_list: D004272:DNA, Mitochondrial; D046911:Macromolecular Substances; D016660:NAD(P)H Dehydrogenase (Quinone); D011808:Quinone Reductases