Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Jul;173(14):4347–4352. doi: 10.1128/jb.173.14.4347-4352.1991

Conjugative transposition of Tn916 requires the excisive and integrative activities of the transposon-encoded integrase.

M J Storrs 1, C Poyart-Salmeron 1, P Trieu-Cuot 1, P Courvalin 1
PMCID: PMC208095  PMID: 1648556

Abstract

Transposon Tn916 is a 16.4-kb broad-host-range conjugative transposon originally detected in the chromosome of Enterococcus faecalis DS16. Transposition of Tn916 and related transposons involves excision of a free, nonreplicative, covalently closed circular intermediate that is substrate for integration. Excisive recombination requires two transposon-encoded proteins, Xis-Tn and Int-Tn, whereas the latter protein alone is sufficient for integration. Here we report that conjugative transposition of Tn916 requires the presence of a functional integrase in both donor and recipient strains. We have constructed a mutant, designated Tn916-int1, by replacing the gene directing synthesis of Int-Tn by an allele inactivated in vitro. In mating experiments, transfer of Tn916-int1 from Bacillus subtilis to E. faecalis was detected only when the transposon-encoded integrase was supplied by trans-complementation in both the donor and the recipient. These results suggest that conjugative transposition of Tn916 requires circularization of the element in the donor followed by transfer and integration of the nonreplicative intermediate in the recipient.

Full text

PDF
4347

Images in this article

4349Image
on p.4349
4350Image
on p.4350

Selected References

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

  1. Appleyard R K. Segregation of New Lysogenic Types during Growth of a Doubly Lysogenic Strain Derived from Escherichia Coli K12. Genetics. 1954 Jul;39(4):440–452. doi: 10.1093/genetics/39.4.440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  3. Brisson-Noël A., Dutka-Malen S., Molinas C., Leclercq R., Courvalin P. Cloning and heterospecific expression of the resistance determinant vanA encoding high-level resistance to glycopeptides in Enterococcus faecium BM4147. Antimicrob Agents Chemother. 1990 May;34(5):924–927. doi: 10.1128/aac.34.5.924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Caillaud F., Carlier C., Courvalin P. Physical analysis of the conjugative shuttle transposon Tn1545. Plasmid. 1987 Jan;17(1):58–60. doi: 10.1016/0147-619x(87)90009-6. [DOI] [PubMed] [Google Scholar]
  5. Caillaud F., Courvalin P. Nucleotide sequence of the ends of the conjugative shuttle transposon Tn1545. Mol Gen Genet. 1987 Aug;209(1):110–115. doi: 10.1007/BF00329844. [DOI] [PubMed] [Google Scholar]
  6. Caparon M. G., Scott J. R. Excision and insertion of the conjugative transposon Tn916 involves a novel recombination mechanism. Cell. 1989 Dec 22;59(6):1027–1034. doi: 10.1016/0092-8674(89)90759-9. [DOI] [PubMed] [Google Scholar]
  7. Christie P. J., Korman R. Z., Zahler S. A., Adsit J. C., Dunny G. M. Two conjugation systems associated with Streptococcus faecalis plasmid pCF10: identification of a conjugative transposon that transfers between S. faecalis and Bacillus subtilis. J Bacteriol. 1987 Jun;169(6):2529–2536. doi: 10.1128/jb.169.6.2529-2536.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clewell D. B., Flannagan S. E., Ike Y., Jones J. M., Gawron-Burke C. Sequence analysis of termini of conjugative transposon Tn916. J Bacteriol. 1988 Jul;170(7):3046–3052. doi: 10.1128/jb.170.7.3046-3052.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clewell D. B., Gawron-Burke C. Conjugative transposons and the dissemination of antibiotic resistance in streptococci. Annu Rev Microbiol. 1986;40:635–659. doi: 10.1146/annurev.mi.40.100186.003223. [DOI] [PubMed] [Google Scholar]
  10. Courvalin P., Carlier C. Tn1545: a conjugative shuttle transposon. Mol Gen Genet. 1987 Feb;206(2):259–264. doi: 10.1007/BF00333582. [DOI] [PubMed] [Google Scholar]
  11. Courvalin P., Carlier C. Transposable multiple antibiotic resistance in Streptococcus pneumoniae. Mol Gen Genet. 1986 Nov;205(2):291–297. doi: 10.1007/BF00430441. [DOI] [PubMed] [Google Scholar]
  12. Ehrlich S. D. DNA cloning in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1433–1436. doi: 10.1073/pnas.75.3.1433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  14. Fitzgerald G. F., Clewell D. B. A conjugative transposon (Tn919) in Streptococcus sanguis. Infect Immun. 1985 Feb;47(2):415–420. doi: 10.1128/iai.47.2.415-420.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fletcher H. M., Marri L., Daneo-Moore L. Transposon-916-like elements in clinical isolates of Enterococcus faecium. J Gen Microbiol. 1989 Nov;135(11):3067–3077. doi: 10.1099/00221287-135-11-3067. [DOI] [PubMed] [Google Scholar]
  16. Franke A. E., Clewell D. B. Evidence for a chromosome-borne resistance transposon (Tn916) in Streptococcus faecalis that is capable of "conjugal" transfer in the absence of a conjugative plasmid. J Bacteriol. 1981 Jan;145(1):494–502. doi: 10.1128/jb.145.1.494-502.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gawron-Burke C., Clewell D. B. A transposon in Streptococcus faecalis with fertility properties. Nature. 1982 Nov 18;300(5889):281–284. doi: 10.1038/300281a0. [DOI] [PubMed] [Google Scholar]
  18. Gawron-Burke C., Clewell D. B. Regeneration of insertionally inactivated streptococcal DNA fragments after excision of transposon Tn916 in Escherichia coli: strategy for targeting and cloning of genes from gram-positive bacteria. J Bacteriol. 1984 Jul;159(1):214–221. doi: 10.1128/jb.159.1.214-221.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hächler H., Kayser F. H., Berger-Bächi B. Homology of a transferable tetracycline resistance determinant of Clostridium difficile with Streptococcus (Enterococcus) faecalis transposon Tn916. Antimicrob Agents Chemother. 1987 Jul;31(7):1033–1038. doi: 10.1128/aac.31.7.1033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ionesco H., Michel J., Cami B., Schaeffer P. Symposium on bacterial spores: II. Genetics of sporulation in Bacillus subtilis Marburg. J Appl Bacteriol. 1970 Mar;33(1):13–24. doi: 10.1111/j.1365-2672.1970.tb05230.x. [DOI] [PubMed] [Google Scholar]
  21. Mabilat C., Goussard S., Sougakoff W., Spencer R. C., Courvalin P. Direct sequencing of the amplified structural gene and promoter for the extended-broad-spectrum beta-lactamase TEM-9 (RHH-1) of Klebsiella pneumoniae. Plasmid. 1990 Jan;23(1):27–34. doi: 10.1016/0147-619x(90)90041-a. [DOI] [PubMed] [Google Scholar]
  22. Naglich J. G., Andrews R. E., Jr Tn916-dependent conjugal transfer of PC194 and PUB110 from Bacillus subtilis into Bacillus thuringiensis subsp. israelensis. Plasmid. 1988 Sep;20(2):113–126. doi: 10.1016/0147-619x(88)90014-5. [DOI] [PubMed] [Google Scholar]
  23. Poyart-Salmeron C., Trieu-Cuot P., Carlier C., Courvalin P. Molecular characterization of two proteins involved in the excision of the conjugative transposon Tn1545: homologies with other site-specific recombinases. EMBO J. 1989 Aug;8(8):2425–2433. doi: 10.1002/j.1460-2075.1989.tb08373.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Poyart-Salmeron C., Trieu-Cuot P., Carlier C., Courvalin P. The integration-excision system of the conjugative transposon Tn 1545 is structurally and functionally related to those of lambdoid phages. Mol Microbiol. 1990 Sep;4(9):1513–1521. doi: 10.1111/j.1365-2958.1990.tb02062.x. [DOI] [PubMed] [Google Scholar]
  25. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  26. Scott J. R., Kirchman P. A., Caparon M. G. An intermediate in transposition of the conjugative transposon Tn916. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4809–4813. doi: 10.1073/pnas.85.13.4809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Senghas E., Jones J. M., Yamamoto M., Gawron-Burke C., Clewell D. B. Genetic organization of the bacterial conjugative transposon Tn916. J Bacteriol. 1988 Jan;170(1):245–249. doi: 10.1128/jb.170.1.245-249.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Spratt B. G., Hedge P. J., te Heesen S., Edelman A., Broome-Smith J. K. Kanamycin-resistant vectors that are analogues of plasmids pUC8, pUC9, pEMBL8 and pEMBL9. Gene. 1986;41(2-3):337–342. doi: 10.1016/0378-1119(86)90117-4. [DOI] [PubMed] [Google Scholar]
  29. Thomas C. M., Smith C. A. Incompatibility group P plasmids: genetics, evolution, and use in genetic manipulation. Annu Rev Microbiol. 1987;41:77–101. doi: 10.1146/annurev.mi.41.100187.000453. [DOI] [PubMed] [Google Scholar]
  30. Torres O. R., Korman R. Z., Zahler S. A., Dunny G. M. The conjugative transposon Tn925: enhancement of conjugal transfer by tetracycline in Enterococcus faecalis and mobilization of chromosomal genes in Bacillus subtilis and E. faecalis. Mol Gen Genet. 1991 Mar;225(3):395–400. doi: 10.1007/BF00261679. [DOI] [PubMed] [Google Scholar]
  31. Trieu-Cuot P., Poyart-Salmeron C., Carlier C., Courvalin P. Nucleotide sequence of the erythromycin resistance gene of the conjugative transposon Tn1545. Nucleic Acids Res. 1990 Jun 25;18(12):3660–3660. doi: 10.1093/nar/18.12.3660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Willetts N., Wilkins B. Processing of plasmid DNA during bacterial conjugation. Microbiol Rev. 1984 Mar;48(1):24–41. doi: 10.1128/mr.48.1.24-41.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yansura D. G., Henner D. J. Use of the Escherichia coli lac repressor and operator to control gene expression in Bacillus subtilis. Proc Natl Acad Sci U S A. 1984 Jan;81(2):439–443. doi: 10.1073/pnas.81.2.439. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES