Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1987 Sep;6(9):2849–2853. doi: 10.1002/j.1460-2075.1987.tb02582.x

Construction of hybrid Tn501/Tn21 transposases in vivo: identification of a region of transposase conferring specificity of recognition of the 38-bp terminal inverted repeats.

L R Evans 1, N L Brown 1
PMCID: PMC553712  PMID: 2824195

Abstract

In order to study the transposase enzymes of Class II prokaryotic transposable elements, we have constructed genes encoding hybrid transposase proteins. This was done by recombination in vivo between the tnpA genes of transposons Tn501 and Tn21. These hybrid genes can complement in trans a transposition-defective mutant of Tn501. The structures of the products of this complementation indicate whether the specificity of the hybrid transposase in recognising the 38 bp terminal inverted repeats is that of Tn501 or that of Tn21. The determinant of this specificity is in the N-terminal region of the transposase protein, between amino acids 28 and 216. The predicted amino acid sequences so far determined of transposases from the Class II family reveal an area of homology in this region.

Full text

PDF

Images in this article

2851Fig. 2.
on p.2851

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Achtman M., Willetts N., Clark A. J. Beginning a genetic analysis of conjugational transfer determined by the F factor in Escherichia coli by isolation and characterization of transfer-deficient mutants. J Bacteriol. 1971 May;106(2):529–538. doi: 10.1128/jb.106.2.529-538.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett P. M., Richmond M. H. Translocation of a discrete piece of deoxyribonucleic acid carrying an amp gene between replicons in Eschericha coli. J Bacteriol. 1976 Apr;126(1):1–6. doi: 10.1128/jb.126.1.1-6.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown N. L., Choi C. L., Grinsted J., Richmond M. H., Whitehead P. R. Nucleotide sequences at the ends of the mercury resistance transposon, Tn501. Nucleic Acids Res. 1980 May 10;8(9):1933–1945. doi: 10.1093/nar/8.9.1933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown N. L., Winnie J. N., Fritzinger D., Pridmore R. D. The nucleotide sequence of the tnpA gene completes the sequence of the Pseudomonas transposon Tn501. Nucleic Acids Res. 1985 Aug 12;13(15):5657–5669. doi: 10.1093/nar/13.15.5657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Craigie R., Arndt-Jovin D. J., Mizuuchi K. A defined system for the DNA strand-transfer reaction at the initiation of bacteriophage Mu transposition: protein and DNA substrate requirements. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7570–7574. doi: 10.1073/pnas.82.22.7570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Craigie R., Mizuuchi K. Mechanism of transposition of bacteriophage Mu: structure of a transposition intermediate. Cell. 1985 Jul;41(3):867–876. doi: 10.1016/s0092-8674(85)80067-2. [DOI] [PubMed] [Google Scholar]
  8. Datta N., Hedges R. W. Trimethoprim resistance conferred by W plasmids in Enterobacteriaceae. J Gen Microbiol. 1972 Sep;72(2):349–355. doi: 10.1099/00221287-72-2-349. [DOI] [PubMed] [Google Scholar]
  9. Gill R., Heffron F., Dougan G., Falkow S. Analysis of sequences transposed by complementation of two classes of transposition-deficient mutants of Tn3. J Bacteriol. 1978 Nov;136(2):742–756. doi: 10.1128/jb.136.2.742-756.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grindley N. D., Reed R. R. Transpositional recombination in prokaryotes. Annu Rev Biochem. 1985;54:863–896. doi: 10.1146/annurev.bi.54.070185.004243. [DOI] [PubMed] [Google Scholar]
  11. Grinsted J., Brown N. L. A Tn21 terminal sequence within Tn501: complementation of tnpA gene function and transposon evolution. Mol Gen Genet. 1984;197(3):497–502. doi: 10.1007/BF00329949. [DOI] [PubMed] [Google Scholar]
  12. Grinsted J., de la Cruz F., Altenbuchner J., Schmitt R. Complementation of transposition of tnpA mutants of Tn3, Tn21, Tn501, and Tn1721. Plasmid. 1982 Nov;8(3):276–286. doi: 10.1016/0147-619x(82)90065-8. [DOI] [PubMed] [Google Scholar]
  13. Heffron F., McCarthy B. J., Ohtsubo H., Ohtsubo E. DNA sequence analysis of the transposon Tn3: three genes and three sites involved in transposition of Tn3. Cell. 1979 Dec;18(4):1153–1163. doi: 10.1016/0092-8674(79)90228-9. [DOI] [PubMed] [Google Scholar]
  14. Kleckner N. Transposable elements in prokaryotes. Annu Rev Genet. 1981;15:341–404. doi: 10.1146/annurev.ge.15.120181.002013. [DOI] [PubMed] [Google Scholar]
  15. Korneluk R. G., Quan F., Gravel R. A. Rapid and reliable dideoxy sequencing of double-stranded DNA. Gene. 1985;40(2-3):317–323. doi: 10.1016/0378-1119(85)90055-1. [DOI] [PubMed] [Google Scholar]
  16. Lund P. A., Ford S. J., Brown N. L. Transcriptional regulation of the mercury-resistance genes of transposon Tn501. J Gen Microbiol. 1986 Feb;132(2):465–480. doi: 10.1099/00221287-132-2-465. [DOI] [PubMed] [Google Scholar]
  17. Michiels T., Cornelis G., Ellis K., Grinsted J. Tn2501, a component of the lactose transposon Tn951, is an example of a new category of class II transposable elements. J Bacteriol. 1987 Feb;169(2):624–631. doi: 10.1128/jb.169.2.624-631.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murray N. E., Bruce S. A., Murray K. Molecular cloning of the DNA ligase gene from bacteriophage T4. II. Amplification and preparation of the gene product. J Mol Biol. 1979 Aug 15;132(3):493–505. doi: 10.1016/0022-2836(79)90271-7. [DOI] [PubMed] [Google Scholar]
  19. Mötsch S., Schmitt R., Avila P., de la Cruz F., Ward E., Grinsted J. Junction sequences generated by 'one-ended transposition'. Nucleic Acids Res. 1985 May 10;13(9):3335–3342. doi: 10.1093/nar/13.9.3335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pabo C. O., Sauer R. T. Protein-DNA recognition. Annu Rev Biochem. 1984;53:293–321. doi: 10.1146/annurev.bi.53.070184.001453. [DOI] [PubMed] [Google Scholar]
  21. Shapiro J. A. Molecular model for the transposition and replication of bacteriophage Mu and other transposable elements. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1933–1937. doi: 10.1073/pnas.76.4.1933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shaw J. H., Clewell D. B. Complete nucleotide sequence of macrolide-lincosamide-streptogramin B-resistance transposon Tn917 in Streptococcus faecalis. J Bacteriol. 1985 Nov;164(2):782–796. doi: 10.1128/jb.164.2.782-796.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ward E., Grinsted J. The nucleotide sequence of the tnpA gene of Tn21. Nucleic Acids Res. 1987 Feb 25;15(4):1799–1806. doi: 10.1093/nar/15.4.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wishart W. L., Broach J. R., Ohtsubo E. ATP-dependent specific binding of Tn3 transposase to Tn3 inverted repeats. Nature. 1985 Apr 11;314(6011):556–558. doi: 10.1038/314556a0. [DOI] [PubMed] [Google Scholar]
  25. 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]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES