title: The F1F0-ATPase of Escherichia coli. The substitution of alanine by tyrosine at position 25 in the c-subunit affects function but not assembly.
abstract: A site-directed mutation in the gene which codes for the c-subunit of the F1F0-ATPase, resulting in the substitution of Ala-25 by Tyr, has been constructed and characterized. A plasmid carrying the mutation was used to transform strain AN943 (uncE429). The resulting strain is unable to grow on succinate as sole carbon source and possesses an uncoupled growth yield. Membranes prepared from the mutant possess low levels of ATPase activity and are proton-impermeable. The F1-ATPase activity was found to be inhibited by 80% when bound to the membrane. When carried on a plasmid, the mutation is dominant in complementation tests with all mutant unc alleles tested and when transformed into wild-type strain AN346, the mutation results in an uncoupled phenotype. A mutant which overcomes this dominance was isolated and found to possess an 11-amino-acid deletion extending from Ile-55 to Met-65 within the c-subunit. These results are discussed in relation to the previously isolated Ala-25 to Thr mutant (Fimmel, A.L., Jans, D.A., Hatch, L., James, L.B., Gibson, F. and Cox, G.B. (1985) Biochim. Biophys. Acta 808, 252-258) and in relation to a previously proposed model for the F0 (Cox, G.B., Fimmel, A.L., Gibson, F. and Hatch, L. (1986) Biochim. Biophys. Acta 849, 62-69).
authors.affiliation: Department of Biochemistry, Faculty of Science, Australian National University, Canberra.
authors.forename: A L
authors.initials: AL
authors.lastname: Fimmel
authors.forename: S A
authors.initials: SA
authors.lastname: Fordham
mesh_terms: D000409:Alanine; D000483:Alleles; D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology*; D005453:Fluorescence; D009154:Mutation; D006180:Proton-Translocating ATPases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011796:Quinacrine; D013329:Structure-Activity Relationship; D014443:Tyrosine
chemical_list: D014443:Tyrosine; D006180:Proton-Translocating ATPases; D011796:Quinacrine; D000409:Alanine