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. 1995 Dec;61(12):4223–4229. doi: 10.1128/aem.61.12.4223-4229.1995

Structure and function of Tn5467, a Tn21-like transposon located on the Thiobacillus ferrooxidans broad-host-range plasmid pTF-FC2.

A M Clennel 1, B Johnston 1, D E Rawlings 1
PMCID: PMC167733  PMID: 8534089

Abstract

A 3.5-kb region of plasmid pTF-FC2, which contains a transposon-like element designated Tn5467, has been sequenced, and its biological activity has been investigated. The transposon is bordered by two 38-bp inverted repeat sequences which have sequence identity in 37 of 38 and in 38 of 39 bp to the tnpA distal and tnpA proximal inverted repeats of Tn21, respectively. Within these borders, open reading frames with amino acid similarity to a glutaredoxin-like protein, a MerR regulatory protein, and a multidrug-resistant-membrane transport-like protein were found. The gene for the glutaredoxin-like protein was expressed in Escherichia coli and enabled growth of a glutathione-requiring E. coli trxA gshA mutant on minimal medium and the reduction of methionine sulfoxide to methionine. In addition, there were two regions which, when translated, had homology to 85% of the N-terminal region of the Tn21 resolvase (tnpR) and to 15% of the C terminus of the Tn21 transposase (tnpA). A region containing res-like sites was located immediately upstream of the partial tnpR gene. Neither the partial transposase nor the resolvase genes of Tn5467 were biologically active, but Tn5467 was transposed and resolved when the Tn21 transposase and resolvase were provided in trans. Tn5467 appears to be a defective transposon which belongs to the Tn21 subgroup of the Tn3 family.

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Selected References

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  1. Avila P., de la Cruz F., Ward E., Grinsted J. Plasmids containing one inverted repeat of Tn21 can fuse with other plasmids in the presence of Tn21 transposase. Mol Gen Genet. 1984;195(1-2):288–293. doi: 10.1007/BF00332761. [DOI] [PubMed] [Google Scholar]
  2. Christie G. E., White T. J., Goodwin T. S. A merR homologue at 74 minutes on the Escherichia coli genome. Gene. 1994 Aug 19;146(1):131–132. doi: 10.1016/0378-1119(94)90847-8. [DOI] [PubMed] [Google Scholar]
  3. Dorrington R. A., Rawlings D. E. Characterization of the minimum replicon of the broad-host-range plasmid pTF-FC2 and similarity between pTF-FC2 and the IncQ plasmids. J Bacteriol. 1990 Oct;172(10):5697–5705. doi: 10.1128/jb.172.10.5697-5705.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eberl L., Kristensen C. S., Givskov M., Grohmann E., Gerlitz M., Schwab H. Analysis of the multimer resolution system encoded by the parCBA operon of broad-host-range plasmid RP4. Mol Microbiol. 1994 Apr;12(1):131–141. doi: 10.1111/j.1365-2958.1994.tb01002.x. [DOI] [PubMed] [Google Scholar]
  5. Grinsted J., de la Cruz F., Schmitt R. The Tn21 subgroup of bacterial transposable elements. Plasmid. 1990 Nov;24(3):163–189. doi: 10.1016/0147-619x(90)90001-s. [DOI] [PubMed] [Google Scholar]
  6. Herrera G., Snyman S. J., Thomson J. A. Construction of a Bioinsecticidal Strain of Pseudomonas fluorescens Active against the Sugarcane Borer, Eldana saccharina. Appl Environ Microbiol. 1994 Feb;60(2):682–690. doi: 10.1128/aem.60.2.682-690.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hughes V. M., Datta N. Conjugative plasmids in bacteria of the 'pre-antibiotic' era. Nature. 1983 Apr 21;302(5910):725–726. doi: 10.1038/302725a0. [DOI] [PubMed] [Google Scholar]
  8. Inoue C., Sugawara K., Kusano T. The merR regulatory gene in Thiobacillus ferrooxidans is spaced apart from the mer structural genes. Mol Microbiol. 1991 Nov;5(11):2707–2718. doi: 10.1111/j.1365-2958.1991.tb01979.x. [DOI] [PubMed] [Google Scholar]
  9. Lewis K. Multidrug resistance pumps in bacteria: variations on a theme. Trends Biochem Sci. 1994 Mar;19(3):119–123. doi: 10.1016/0968-0004(94)90204-6. [DOI] [PubMed] [Google Scholar]
  10. Lim C. J., Gleason F. K., Fuchs J. A. Cloning, expression, and characterization of the Anabaena thioredoxin gene in Escherichia coli. J Bacteriol. 1986 Dec;168(3):1258–1264. doi: 10.1128/jb.168.3.1258-1264.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. McFarlan S. C., Terrell C. A., Hogenkamp H. P. The purification, characterization, and primary structure of a small redox protein from Methanobacterium thermoautotrophicum, an archaebacterium. J Biol Chem. 1992 May 25;267(15):10561–10569. [PubMed] [Google Scholar]
  12. Rawlings D. E., Kusano T. Molecular genetics of Thiobacillus ferrooxidans. Microbiol Rev. 1994 Mar;58(1):39–55. doi: 10.1128/mr.58.1.39-55.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rawlings D. E., Pretorius I., Woods D. R. Expression of a Thiobacillus ferrooxidans origin of replication in Escherichia coli. J Bacteriol. 1984 May;158(2):737–738. doi: 10.1128/jb.158.2.737-738.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rogowsky P., Halford S. E., Schmitt R. Definition of three resolvase binding sites at the res loci of Tn21 and Tn1721. EMBO J. 1985 Aug;4(8):2135–2141. doi: 10.1002/j.1460-2075.1985.tb03904.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rohrer J., Rawlings D. E. Sequence analysis and characterization of the mobilization region of a broad-host-range plasmid, pTF-FC2, isolated from Thiobacillus ferrooxidans. J Bacteriol. 1992 Oct;174(19):6230–6237. doi: 10.1128/jb.174.19.6230-6237.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shiratori T., Inoue C., Sugawara K., Kusano T., Kitagawa Y. Cloning and expression of Thiobacillus ferrooxidans mercury ion resistance genes in Escherichia coli. J Bacteriol. 1989 Jun;171(6):3458–3464. doi: 10.1128/jb.171.6.3458-3464.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Silver S., Walderhaug M. Gene regulation of plasmid- and chromosome-determined inorganic ion transport in bacteria. Microbiol Rev. 1992 Mar;56(1):195–228. doi: 10.1128/mr.56.1.195-228.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. de la Cruz F., Grinsted J. Genetic and molecular characterization of Tn21, a multiple resistance transposon from R100.1. J Bacteriol. 1982 Jul;151(1):222–228. doi: 10.1128/jb.151.1.222-228.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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