Abstract
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.
Full text
PDF![874](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/369c/215341/2767e008857a/jbacter00238-0186.png)
![875](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/369c/215341/4f6b9a17a06a/jbacter00238-0187.png)
![876](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/369c/215341/5b540a2ecfed/jbacter00238-0188.png)
![877](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/369c/215341/c43a2a92b6cc/jbacter00238-0189.png)
![878](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/369c/215341/bbeb6d0ccbcf/jbacter00238-0190.png)
![879](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/369c/215341/24958b61c709/jbacter00238-0191.png)
![880](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/369c/215341/f7e5fceb2fc0/jbacter00238-0192.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ames G. F. Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974 Jan 25;249(2):634–644. [PubMed] [Google Scholar]
- Bindereif A., Braun V., Hantke K. The cloacin receptor of ColV-bearing Escherichia coli is part of the Fe3+-aerobactin transport system. J Bacteriol. 1982 Jun;150(3):1472–1475. doi: 10.1128/jb.150.3.1472-1475.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bindereif A., Neilands J. B. Cloning of the aerobactin-mediated iron assimilation system of plasmid ColV. J Bacteriol. 1983 Feb;153(2):1111–1113. doi: 10.1128/jb.153.2.1111-1113.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clarke L., Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. doi: 10.1016/0092-8674(76)90055-6. [DOI] [PubMed] [Google Scholar]
- Cronan J. E., Jr Beta-alanine synthesis in Escherichia coli. J Bacteriol. 1980 Mar;141(3):1291–1297. doi: 10.1128/jb.141.3.1291-1297.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Csonka L. N., Clark A. J. Construction of an Hfr strain useful for transferring recA mutations between Escherichia coli strains. J Bacteriol. 1980 Jul;143(1):529–530. doi: 10.1128/jb.143.1.529-530.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guterman S. K. Colicin B: mode of action and inhibition by enterochelin. J Bacteriol. 1973 Jun;114(3):1217–1224. doi: 10.1128/jb.114.3.1217-1224.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hancock R. E., Hantke K., Braun V. Iron transport of Escherichia coli K-12: involvement of the colicin B receptor and of a citrate-inducible protein. J Bacteriol. 1976 Sep;127(3):1370–1375. doi: 10.1128/jb.127.3.1370-1375.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hartmann A., Braun V. Iron transport in Escherichia coli: uptake and modification of ferrichrome. J Bacteriol. 1980 Jul;143(1):246–255. doi: 10.1128/jb.143.1.246-255.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
- Jorgensen R. A., Rothstein S. J., Reznikoff W. S. A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. Mol Gen Genet. 1979;177(1):65–72. doi: 10.1007/BF00267254. [DOI] [PubMed] [Google Scholar]
- Kadner R. J., Heller K., Coulton J. W., Braun V. Genetic control of hydroxamate-mediated iron uptake in Escherichia coli. J Bacteriol. 1980 Jul;143(1):256–264. doi: 10.1128/jb.143.1.256-264.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Katz L., Kingsbury D. T., Helinski D. R. Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex. J Bacteriol. 1973 May;114(2):577–591. doi: 10.1128/jb.114.2.577-591.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leong J., Neilands J. B. Mechanisms of siderophore iron transport in enteric bacteria. J Bacteriol. 1976 May;126(2):823–830. doi: 10.1128/jb.126.2.823-830.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Linke W. D., Crueger A., Diekmann H. Stoffwechselprodukte von Miktoorganismen. 106. Zur Konstitution des Terregens-Faktors. Arch Mikrobiol. 1972;85(1):44–50. [PubMed] [Google Scholar]
- Llinás M., Neilands J. B. The structure of two alanine containing ferrichromes: sequence determination by proton magnetic resonance. Biophys Struct Mech. 1976 Aug 23;2(2):105–117. doi: 10.1007/BF00863704. [DOI] [PubMed] [Google Scholar]
- Low B. Formation of merodiploids in matings with a class of Rec- recipient strains of Escherichia coli K12. Proc Natl Acad Sci U S A. 1968 May;60(1):160–167. doi: 10.1073/pnas.60.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Luckey M., Pollack J. R., Wayne R., Ames B. N., Neilands J. B. Iron uptake in Salmonella typhimurium: utilization of exogenous siderochromes as iron carriers. J Bacteriol. 1972 Sep;111(3):731–738. doi: 10.1128/jb.111.3.731-738.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
- Negrin R. S., Neilands J. B. Ferrichrome transport in inner membrane vesicles of Escherichia coli K12. J Biol Chem. 1978 Apr 10;253(7):2339–2342. [PubMed] [Google Scholar]
- Neilands J. B. Microbial envelope proteins related to iron. Annu Rev Microbiol. 1982;36:285–309. doi: 10.1146/annurev.mi.36.100182.001441. [DOI] [PubMed] [Google Scholar]
- Plastow G. S., Pratt J. M., Holland I. B. The ferrichrome receptor protein (tonA) of Escherichia coli is synthesised as a precursor in vitro. FEBS Lett. 1981 Aug 31;131(2):262–264. doi: 10.1016/0014-5793(81)80380-8. [DOI] [PubMed] [Google Scholar]
- Ramos S., Kaback H. R. The relationship between the electrochemical proton gradient and active transport in Escherichia coli membrane vesicles. Biochemistry. 1977 Mar 8;16(5):854–859. doi: 10.1021/bi00624a007. [DOI] [PubMed] [Google Scholar]
- Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sancar A., Wharton R. P., Seltzer S., Kacinski B. M., Clarke N. D., Rupp W. D. Identification of the uvrA gene product. J Mol Biol. 1981 May 5;148(1):45–62. doi: 10.1016/0022-2836(81)90234-5. [DOI] [PubMed] [Google Scholar]
- Takeda Y., Nishimura A., Nishimura Y., Yamada M., Yasuda S., Suzuki H., Hirota Y. Synthetic ColE1 Plasmids carrying genes for penicillin-binding proteins in Escherichia coli. Plasmid. 1981 Jul;6(1):86–98. doi: 10.1016/0147-619x(81)90056-1. [DOI] [PubMed] [Google Scholar]
- Thummel C. S., Burgess T. L., Tjian R. Properties of simian virus 40 small t antigen overproduced in bacteria. J Virol. 1981 Feb;37(2):683–697. doi: 10.1128/jvi.37.2.683-697.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wayne R., Frick K., Neilands J. B. Siderophore protection against colicins M, B, V, and Ia in Escherichia coli. J Bacteriol. 1976 Apr;126(1):7–12. doi: 10.1128/jb.126.1.7-12.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wayne R., Neilands J. B. Evidence for common binding sites for ferrichrome compounds and bacteriophage phi 80 in the cell envelope of Escherichia coli. J Bacteriol. 1975 Feb;121(2):497–503. doi: 10.1128/jb.121.2.497-503.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weaver C. A., Konisky J. tonB-independent ferrichrome-mediated iron transport in Escherichia coli spheroplasts. J Bacteriol. 1980 Sep;143(3):1513–1518. doi: 10.1128/jb.143.3.1513-1518.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wookey P. J., Hussein S., Braun V. Functions in outer and inner membranes of Escherichia coli for ferrichrome transport. J Bacteriol. 1981 Jun;146(3):1158–1161. doi: 10.1128/jb.146.3.1158-1161.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zehnbauer B. A., Markovitz A. Cloning of gene lon (capR) of Escherichia coli K-12 and identification of polypeptides specified by the cloned deoxyribonucleic acid fragment. J Bacteriol. 1980 Aug;143(2):852–863. doi: 10.1128/jb.143.2.852-863.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Boer H. A., Comstock L. J., Vasser M. The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci U S A. 1983 Jan;80(1):21–25. doi: 10.1073/pnas.80.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]