The genes for the eight subunits of the membrane bound ATP synthase of Escherichia coli

Abstract

The genes for the eight subunits of the membrane bound ATP synthase of Escherichia coli (Ca++, Mg++ dependent ATPase, EC 3.6.1.3) were mapped through genetic, physical and functional analysis of specialized transducing phages lambda asn (von Meyenburg et al. 1978). The ATP synthase genes, designated atp1, are located at 83.2 min in a segment of the chromosome between 3.5 and 11.3 kb left (counterclockwise) of the origin of replication oriC. The counterclockwise order of the genes for the eight subunits, the expression of which starts from a control region at 3.5 kb-L, was found to be: a, (c, b, delta), alpha, gamma, (epsilon, beta) which in the notation of Downie el al. (1981) reads atp B (EFH) A G (C D). The analysis was in part based on the isolation of new types of atp (unc, Suc-) mutations. We made use of the fact that specialized transducing phages lambda asn carrying oriC can establish themselves as minichromosomes rendering asnA cells Asn+, and that the resulting Asn+ cells grow slowly if the lambda asn carries part or all of the atp operon. Selecting for fast growing strains mutations were isolated on the lambda asn which either eliminated atp genes or affected their expression ("promoter" mutations). The relationship between these atp mutations and the cop mutations of Ogura et al. (1980), which also appear to map in front of or within the atp genes, is discussed.

The proton conducting F0-part of bacterial ATP synthases

“…As described in subseetion lIlA, the large polar domain of subunit b is easily attacked by proteinases in vitro [49]. Another deletion mutant CM 1470 has a large deletion including genes of subunits a, C, b, 0 and part of a [13]. The membrane of this strain is devoid of all Fa subunits.…”

“…As described in section II (Fig. 1) and reviewed in detail elsewhere, the E. eoli F o F 1 is coded for by an operon of eight genes [1,12,13]. The genes of F o preceding the F 1 genes are transcribed in the order a, e, b.…”

“…Accord- [8,10] and analyzed by SDSjpolyacrylamide gel eleetrophoresis. The order of the genes in the une operon and the eorrelation of genes with A TP-synthase subunits was established genetieally and by nucleotide sequenee analysis [1,[12][13][14][15][16][17][18][19][20][21][22][23]. The numbers below the genes indieate the number of amino acid residues in the subunit polypeptide.…”

“…The whole ATP synthase contains, in addition, the five F I subunits IX, ß, y, 8 and E. These eight subunit polypeptides are coded for by the eight genes that form the alp (une) operon (see section IV). The alp operon has been weIl defined geneticaIly, especially by Downie, et al [1,12], and by Von Meyenburg and co-workers [13], and its nucleotide sequence has been determined [14][15][16][17][18][19][20][21][22][23]. Therefore, the subunit composition of the ATP synthase and F o from E. eoli is proven.…”

“…The genes have been designated une A to une H 01' alp A to alp H by different laboratories. In addition, inhibitor-resistant strains of E. eoli are known which all are affected in the alp E (une E) gene, coding for subunit e. In yeast, the mitochondrial c=J, coding region for the subunits a, e, b; --, indicates the cloned region of the alp operon in the respective plasmid [l01,102]; ------, indicates the deleted region in the two strains [13].…”