cDNA of the 24 kDa subunit of the bovine respiratory chain NADH dehydrogenase: high sequence conservation in mammals and tissue-specific and growth-dependent expression.

We have isolated and sequenced several overlapping cDNA clones from a bovine lambda gt10 library which encode all but the first five amino acids of the entire mature 24 kDa subunit of NADH:ubiquinone oxidoreductase (EC 1.6.99.3), the first enzyme of the respiratory chain. The derived amino acid sequence agrees with that determined by direct sequencing of the purified protein, filling in a gap in the published sequence. A comparison of the nucleotide and amino acid sequences of the bovine 24 kDa subunit with those recently determined for the rat homologue has shown that this nuclear-encoded subunit of an OX-PHOS complex has diverged in these two species much less than the mitochondrial DNA-encoded subunits of the same enzyme complex, and also less than a set of available non-mitochondrial nuclear DNA-coded proteins. The sequence analysis of the clones has revealed the expression in the brain of two mRNAs differing in the length of the 3'-untranslated region. Furthermore, two polyadenylated RNA species, 930 and 1080 nucleotides in length, probably corresponding to the above mRNAs, have been detected in bovine brain and other tissues by RNA gel blot hybridization. The level of expression of the 24kDa subunit gene varies by more than an order of magnitude among different tissues. A cross-hybridizing mRNA species of 930 nucleotides has also been observed in HeLa cells and found to be strongly growth regulated.