Nucleotide sequence of the promoter region of the gene cluster for proton-translocating ATPase from Escherichiacoli and identification of the active promoter (original) (raw)
Related papers
Archives of Biochemistry and Biophysics, 1983
Various hybrid plasmids carrying a portion of the gene for the y subunit of the H'-ATPase of Escherichia coli complemented five mutants defective in the enzyme in a genetic test, indicating that the mutants are defective in the y subunit. Since the nucleotide sequence of genomic DNA carried on the plasmids is known, the defective site(s) of the mutants could be located within the gene for the y subunit as follows: KFlO and NR70, KFl, and KF12 and KF13 have a mutation causing a defect(s) in amino acid residues 1 to 82, 83 to 167, and 168 to 287, respectively, of the y subunit. The biochemical properties of all these mutants except NR70 were analyzed in terms of proton permeability of the membranes and assembly of F1. Results suggested that KFl and KFlO have defective F1 without at least the cy and p subunits on their membranes, whereas KF12 and KF13 have F1's of rather similar structure to that of the wild type. Attempts were made to purify F1 of KF12 as a single complex. Although the F1 complex dissociated during purification, active CY and /3 subunits of KF12 were partially purified. On the basis of these biochemical and genetic results, it is suggested that structural alterations in the primary sequence of the y subunit corresponding to residues 1 to 167 cause more extensive defects in the assembly of F1 than alteration in the sequence of residues 168 to 287. The proton-translocating ATPase of E.scherichia coli catalyzes the hydrolysis and synthesis of ATP reversibly. The enzyme is composed of two distinct portions, F1 and FO," which are extrinsic and intrinsic portions, respectively, of the membranes. F1 has catalytic activity and has five different subunits, CX, p, y, 6, and t (l
Biochemical and Biophysical Research Communications, 1981
The recombinant plasmid pMCR533 carries part of the gene cluster coding for the proton-translocating ATPase (FI-F0) of E. coli. The DNA sequence of the structural gene for dicyclohexylcarbodiimide-binding protein was obtained by analyzing a DNA fragment from this plasmid. The DNA sequence corresponding to the amino terminus of the structural gene for the a subunit was also found on the plasmid. Both sequences were in good agreement with the amino acid sequences determined previously. From the exact locations of these two genes we have located genes coding for other components of FI-F 0 on the defined portion of E. coli DNA. A proposal is made to use the nomenclature pap (proton-translocating ATPase subunit protein) for the structural gene cluster for Fz-F0.
1984
A hybrid plasmid, pKY159, carrying the promoter and the proximal region of the gene cluster for protontranslocating ATPase caused growth inhibition of Escherichia coli cells (K. Yamaguchi and M. Yamaguchi, J. Bacteriol. 153:550-554, 1983). The mechanism of this growth inhibition was studied, especially in terms of the responsible gene(s). Insertion of IS], IS5, or y8 between the promoter and the gene for a possible component of the ATPase of 14,000 daltons (14K protein) released the inhibitory effect by pKY159. Deletion of the gene for subunit a also released the effect. However, deletion in the gene for the 14K protein released the effect only with an additional insertion within the gene. These results suggested that overproduction of subunit a is closely related to growth inhibition, whereas the 14K protein is not. Proton-translocating ATPase (H+-ATPase) has a key function in energy transduction in cells (7, 10-12, 39). The enzyme consists of two distinct portions, F1 and Fo. F1 is the membrane-peripheral portion and has five different subunits, a, Iy, 8, and e. Fo is the membrane-intrinsic portion and has three subunits, a, b, and c. F1 and Fo act as the ATPase and proton channel, respectively, and membranes without
Journal of bacteriology, 1984
A hybrid plasmid, pKY159, carrying the promoter and the proximal region of the gene cluster for proton-translocating ATPase caused growth inhibition of Escherichia coli cells (K. Yamaguchi and M. Yamaguchi, J. Bacteriol. 153:550-554, 1983). The mechanism of this growth inhibition was studied, especially in terms of the responsible gene(s). Insertion of IS1, IS5, or gamma delta between the promoter and the gene for a possible component of the ATPase of 14,000 daltons (14K protein) released the inhibitory effect by pKY159. Deletion of the gene for subunit a also released the effect. However, deletion in the gene for the 14K protein released the effect only with an additional insertion within the gene. These results suggested that overproduction of subunit a is closely related to growth inhibition, whereas the 14K protein is not.
Proceedings of the National Academy of Sciences, 1980
The proton-translocating ATPase (FI-Fo) of oxidative phosphorylation (ATP phosphohydrolase, EC 3.6.1.3) is coded for by a set of structural genes comprising the une operon in Escherichia coli. We have analyzed several new transducing phages and plasmids carrying various lengths of the DNA segments of the unc operon by complementation assay using 14 new unc-mutants and representatives-of previously described strains which were made available to us. Transducing phages carrying parts of the unc gene cluster were isolated: XuncA-9 and Xg mS phages converted only some of the uncmutants to Unc+, as determined by complementation assays. A new hybrid plasmid (pMCR533) carrying part of the unc operon was constructed by inserting the HindIII fragment of Xasn-5 DNA (a phage carrying the entire unc operon) into the unique HindIll site of pBR322. This plasmid transformed eight uncstrains to Unc+, including uncB4O2 and uncA401, but did not complement uncDll or four other strains. Two minichromosomes which carry the E. coli replication origin were also tested: plasmid pNHO5 transformed the uncB402 but not the uncA4Ol strain to Unc+, whereas plasmid pMCFI transformed none of the mutants tested. Analysis of the DNAs from these transducing phages and plasmids with restriction endonucleases suggested that all of the structural genes for the F1-Fo complex are localized within a DNA segment of approximately 4.5 megadaltons containing two EcoRI sites. The approximate locations of the uncmutations were mapped on this DNA segment.