Overproduction of truncated subunit a of H+-ATPase causes growth inhibition of Escherichia coli.
Genes (uncB) for wild-type and mutant a subunits of Escherichia coli H+-ATPase (F0F1) were cloned into recombinant plasmids. The subunits were expressed under the control of a weak promoter of the unc operon at 30 degrees C and strong promoters of lambda phage at 42 degrees C. At 30 degrees C, the wild type and a truncated (Glu-269----end) a subunit complemented the defect of the a subunit mutant KF24A (Trp-111----end), whereas the other mutant subunits (Trp-111----end, Trp-231----end, Gln-252----end, and a subunit with a deletion of residues 21 to 227) did not. Three mutant subunits (Trp-231----end, Gln-252----end, and Glu-269----end) and the wild-type a subunit caused growth inhibition associated with cell elongation, an uneven distribution of membrane proteins, and an altered septum structure when they were expressed at 42 degrees C. These phenomena were not observed with the other mutant subunits, suggesting that overproduction of the middle region (between residues 111 and 230) of the a subunit causes growth inhibition.
Eya
S
S
Department of Organic Chemistry and Biochemistry, Osaka University, Japan.
Maeda
M
M
Tomochika
K
K
Kanemasa
Y
Y
Futai
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004790:Enzyme Induction; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000648:ultrastructure; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D007700:Kinetics; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000032:analysis; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D006180:Proton-Translocating ATPases / Q000096:biosynthesis / Q000235:genetics*; D015183:Restriction Mapping
D046911:Macromolecular Substances; D008565:Membrane Proteins; D006180:Proton-Translocating ATPases