Genetic and biochemical characterization of the Escherichia coli K-12 fhuB mutation.
The fhuB region of Escherichia coli K-12 was subcloned from pLC4-44 into pP lac to obtain pCPN1. Deletions of this recombinant plasmid were made, and a 1.4-kilobase PstI fragment was further subcloned into the vector plasmid pKK177-2 to obtain pCPN12. The response of tonA and tonB strains and fhuB strains containing the plasmids to 15 hydroxamate siderophores were assayed. Results showed that tonA strains were deficient only in the utilization of ferrichrome-type siderophores, whereas fhuB strains were deficient in the utilization of all hydroxamate-type siderophores. The response of the plasmid-containing fhuB strains to the siderophores showed that the fhuB gene resides on a 1.4-kilobase PstI fragment of DNA. The proteins synthesized by these plasmids were examined in maxicells of strain CSR603. Plasmid pCPN1 expressed five proteins of molecular weights 78,000, 40,000, 30,000, 24,000, and 13,700. By the use of deletions of pCPN1, the approximate order of the genes for these proteins was determined. Plasmid pCPN12 expressed no proteins other than the beta-lactamase proteins in maxicell strain CSR603. However, in maxicell strain BN660, a lon mutant, it expressed a 20,000-molecular-weight protein. Inner membrane vesicles made from tonB and fhuB strains were able to transport [55Fe]ferrichrome and [55Fe]rhodotorulate at rates similar to those obtained in vesicles from tonB+ and fhuB+ strains.
D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology; D001692:Biological Transport; D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005291:Ferrichrome / Q000378:metabolism; D005798:Genes, Bacterial; D006877:Hydroxamic Acids / Q000378:metabolism*; D007425:Intracellular Membranes / Q000378:metabolism; D007502:Iron Chelating Agents / Q000378:metabolism*; D009154:Mutation; D010879:Piperazines / Q000378:metabolism; D010957:Plasmids; D017262:Siderophores
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D006877:Hydroxamic Acids; D007502:Iron Chelating Agents; D010879:Piperazines; D017262:Siderophores; D005291:Ferrichrome; C003305:rhodotorulic acid