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1.
Biochim Biophys Acta ; 1089(3): 389-90, 1991 Jul 23.
Article in English | MEDLINE | ID: mdl-1830489

ABSTRACT

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.


Subject(s)
Mitochondria/enzymology , Neurospora crassa/genetics , Quinone Reductases/genetics , Amino Acid Sequence , Base Sequence , Blotting, Northern , DNA, Fungal , Molecular Sequence Data , NAD(P)H Dehydrogenase (Quinone) , Neurospora crassa/enzymology
2.
Biochim Biophys Acta ; 1088(2): 325-6, 1991 Feb 16.
Article in English | MEDLINE | ID: mdl-1825789

ABSTRACT

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.


Subject(s)
DNA, Fungal/genetics , Neurospora crassa/genetics , Quinone Reductases/genetics , Amino Acid Sequence , Base Sequence , Macromolecular Substances , Molecular Sequence Data , Molecular Weight , NAD(P)H Dehydrogenase (Quinone) , Neurospora crassa/enzymology
3.
FEBS Lett ; 278(1): 75-8, 1991 Jan 14.
Article in English | MEDLINE | ID: mdl-1825202

ABSTRACT

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.


Subject(s)
Cytochrome Reductases/genetics , Endopeptidases/genetics , Mitochondria/enzymology , NADH Dehydrogenase/genetics , Amino Acid Sequence , DNA/genetics , Introns , Molecular Sequence Data , Neurospora crassa , Restriction Mapping , Sequence Homology, Nucleic Acid
4.
Curr Genet ; 18(1): 59-64, 1990 Jul.
Article in English | MEDLINE | ID: mdl-2147127

ABSTRACT

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.


Subject(s)
Neurospora crassa/genetics , Quinone Reductases/genetics , Amino Acid Sequence , Base Sequence , Genes, Fungal , Molecular Sequence Data , NAD(P)H Dehydrogenase (Quinone) , Neurospora crassa/enzymology , Open Reading Frames , Quinone Reductases/metabolism , Restriction Mapping , Sequence Homology, Nucleic Acid
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