Mammalian H+-ATP synthase is a supramolecule composed of at least 14 subunits that have a constant stoichiometry. Nevertheless the coordinate regulation of the gene expressions of various subunits remains obscure. To clarify the coordinate transcriptional regulatory system of mammalian H+-ATP synthase, we determined the absolute amount of nine species of mRNAs for eight nuclear-encoded subunits of HC-ATP synthase in different tissues of 8-week-old rats by use of the synthetic mRNAs and P-32-labeled DNA probes for each mRNA. Our quantitative analyses of the transcripts of H+-ATP synthase revealed that nine species of the subunits in different tissues of 8-week-old rats were divisible into two groups: a high transcript gene (HTG) group (beta-subunit, subunit b, subunit d, subunit e, and Factor 6) and a low transcript gene (LTG) group (subunit c(P1), subunit c(P2), IF1, and oligomycin sensitivity-conferring protein). The transcription step of LTG could constitute a bottleneck in the biogenesis of H+-ATP synthase. Thus, the transcriptional regulatory system of the LTG may play a hey role in the biogenesis of mammalian H+-ATP synthase. The HTG were transcribed in a tissue-specific manner that corresponds with energy demand in the tissues. However, there was no tissue specificity in subunit c(P2), Furthermore, the tissue specificity of the transcript of IF1 differed substantially from that of HTG, suggesting that it could be crucial in the protection of mitochondrial membrane under abnormal conditions.