RESUMO
We isolated and sequenced cDNA for the 29.9 kDa subunit of mitochondrial NADH: ubiquinone reductase (complex I) from a Neurospora crassa library in the lambda gt11 expression vector. The N-terminus of the mature protein was determined by Edman-degradation. The cDNA contains an open reading frame encoding a preprotein of 273 amino acids. The presequence of the transit protein essential for mitochondrial import is eight residues long. Northern-blot analysis shows, that the level of the corresponding mRNA is increased 3-fold if cells are grown in the presence of chloramphenicol.
Assuntos
Mitocôndrias/enzimologia , Neurospora crassa/genética , Quinona Redutases/genética , Sequência de Aminoácidos , Sequência de Bases , Northern Blotting , DNA Fúngico , Dados de Sequência Molecular , NAD(P)H Desidrogenase (Quinona) , Neurospora crassa/enzimologiaRESUMO
The primary structure of a nuclear-encoded subunit of the respiratory chain NADH:ubiquinone reductase (complex I) from Neurospora crassa was determined by sequencing cDNA, genomic DNA and the N-terminus of the protein. The sequence correlates to a protein of 200 amino acids and a molecular mass of 21349 Da. The protein is synthesized without a cleavable presequence. It contains two alpha-helices predicted to traverse the bilayer and is a constituent of the membrane part of complex I.
Assuntos
DNA Fúngico/genética , Neurospora crassa/genética , Quinona Redutases/genética , Sequência de Aminoácidos , Sequência de Bases , Substâncias Macromoleculares , Dados de Sequência Molecular , Peso Molecular , NAD(P)H Desidrogenase (Quinona) , Neurospora crassa/enzimologiaRESUMO
The primary structure of a 40 kDa subunit of the respiratory chain NADH:ubiquinone reductase from Neurospora crassa was determined by sequencing cDNA, genomic DNA and the N-terminus of the mature protein. The gene which is interrupted by 7 introns encodes a preprotein consisting of 375 amino acids with a 26 amino acid long presequence typical for a mitochondrial targeting signal. The sequence of the mature subunit shows conspicuous similarities to the recently [(1989) Nature 339, 147-149] discovered protein family which includes subunits I and II of the ubiquinol:cytochrome c reductase, and the processing proteins, matrix processing peptidase and processing enhancing protein, of mitochondria. The possible role of the subunit is discussed.
Assuntos
Redutases do Citocromo/genética , Endopeptidases/genética , Mitocôndrias/enzimologia , NADH Desidrogenase/genética , Sequência de Aminoácidos , DNA/genética , Íntrons , Dados de Sequência Molecular , Neurospora crassa , Mapeamento por Restrição , Homologia de Sequência do Ácido NucleicoRESUMO
The primary structure of the 49 K subunit of the respiratory chain NADH:ubiquinone reductase (complex I) from Neurospora crassa was determined by sequencing cDNA, genomic DNA and the N-terminus of the mature protein. The sequence lengths correlate to a molecular mass of 54,002 daltons for the preprotein and 49,239 daltons for the mature protein. The presequence consists of 42 amino acids of typical composition for sequences which target nuclear-encoded proteins into mitochondria. The mature protein consists of 436 amino acids and shows 64% similarity to a 49 K subunit of bovine heart NADH:ubiquinone reductase and 33% to a predicted translation product of an open reading frame in the chloroplast DNAs of Marchantia polymorpha and Nicotiana tabacum. Evidence for an iron-sulfur cluster in the subunit is discussed.