Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Nov;177(22):6644–6651. doi: 10.1128/jb.177.22.6644-6651.1995

A functional origin of transfer (oriT) on the conjugative transposon Tn916.

D D Jaworski 1, D B Clewell 1
PMCID: PMC177520  PMID: 7592445

Abstract

The origin of transfer (oriT) of the 18-kb conjugative transposon Tn916 has been localized to a 466-bp region which spans nucleotides 15215 to 15681 on the transposon map. The oriT lies within an intercistronic region between open reading frames ORF20 and ORF21 that contains six sets of inverted repeats ranging from 10 to 20 bp in size. The segment contains three sequences showing identity in 9 of 12 bp to the consensus nicking site (nic) of the IncP family of conjugative plasmids found in gram-negative bacteria. Overlapping one of these sequences is a region similar to the nic site of the F plasmid. Functionality was based on the ability of the oriT-containing sequence to provide a cis-acting mobilization of chimeras involving the shuttle vector pWM401 in response to activation in trans by an intact chromosome-borne transposon Tn916 delta E. Cloned segments of 466 or 376 nucleotides resulted in unselected cotransfer of the plasmid at levels of about 40% when selection was for Tn916 delta E, whereas a 110-bp segment resulted in cotransfer at a frequency of about 7%. Mobilization was specific in that gram-positive plasmids, such as pAD1 and pAM beta 1, and the gram-negative plasmids pOX38 (a derivative of F) and RP1 did not mobilize oriT-containing chimeras.

Full Text

The Full Text of this article is available as a PDF (261.5 KB).

Selected References

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

  1. Boyd A. C., Archer J. A., Sherratt D. J. Characterization of the ColE1 mobilization region and its protein products. Mol Gen Genet. 1989 Jun;217(2-3):488–498. doi: 10.1007/BF02464922. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. 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]
  5. Clewell D. B., Flannagan S. E., Jaworski D. D. Unconstrained bacterial promiscuity: the Tn916-Tn1545 family of conjugative transposons. Trends Microbiol. 1995 Jun;3(6):229–236. doi: 10.1016/s0966-842x(00)88930-1. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Clewell D. B. Movable genetic elements and antibiotic resistance in enterococci. Eur J Clin Microbiol Infect Dis. 1990 Feb;9(2):90–102. doi: 10.1007/BF01963632. [DOI] [PubMed] [Google Scholar]
  8. Clewell D. B., Tomich P. K., Gawron-Burke M. C., Franke A. E., Yagi Y., An F. Y. Mapping of Streptococcus faecalis plasmids pAD1 and pAD2 and studies relating to transposition of Tn917. J Bacteriol. 1982 Dec;152(3):1220–1230. doi: 10.1128/jb.152.3.1220-1230.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Flannagan S. E., Clewell D. B. Conjugative transfer of Tn916 in Enterococcus faecalis: trans activation of homologous transposons. J Bacteriol. 1991 Nov;173(22):7136–7141. doi: 10.1128/jb.173.22.7136-7141.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Flannagan S. E., Zitzow L. A., Su Y. A., Clewell D. B. Nucleotide sequence of the 18-kb conjugative transposon Tn916 from Enterococcus faecalis. Plasmid. 1994 Nov;32(3):350–354. doi: 10.1006/plas.1994.1077. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. 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]
  15. Ike Y., Clewell D. B. Genetic analysis of the pAD1 pheromone response in Streptococcus faecalis, using transposon Tn917 as an insertional mutagen. J Bacteriol. 1984 Jun;158(3):777–783. doi: 10.1128/jb.158.3.777-783.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jaworski D. D., Clewell D. B. Evidence that coupling sequences play a frequency-determining role in conjugative transposition of Tn916 in Enterococcus faecalis. J Bacteriol. 1994 Jun;176(11):3328–3335. doi: 10.1128/jb.176.11.3328-3335.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Li L. Y., Shoemaker N. B., Salyers A. A. Location and characteristics of the transfer region of a Bacteroides conjugative transposon and regulation of transfer genes. J Bacteriol. 1995 Sep;177(17):4992–4999. doi: 10.1128/jb.177.17.4992-4999.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lu F., Churchward G. Conjugative transposition: Tn916 integrase contains two independent DNA binding domains that recognize different DNA sequences. EMBO J. 1994 Apr 1;13(7):1541–1548. doi: 10.1002/j.1460-2075.1994.tb06416.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lu F., Churchward G. Tn916 target DNA sequences bind the C-terminal domain of integrase protein with different affinities that correlate with transposon insertion frequency. J Bacteriol. 1995 Apr;177(8):1938–1946. doi: 10.1128/jb.177.8.1938-1946.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pansegrau W., Lanka E. Common sequence motifs in DNA relaxases and nick regions from a variety of DNA transfer systems. Nucleic Acids Res. 1991 Jun 25;19(12):3455–3455. doi: 10.1093/nar/19.12.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rice L. B., Marshall S. H., Carias L. L. Tn5381, a conjugative transposon identifiable as a circular form in Enterococcus faecalis. J Bacteriol. 1992 Nov;174(22):7308–7315. doi: 10.1128/jb.174.22.7308-7315.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rubens C. E., Heggen L. M. Tn916 delta E: a Tn916 transposon derivative expressing erythromycin resistance. Plasmid. 1988 Sep;20(2):137–142. doi: 10.1016/0147-619x(88)90016-9. [DOI] [PubMed] [Google Scholar]
  23. Scott J. R., Bringel F., Marra D., Van Alstine G., Rudy C. K. Conjugative transposition of Tn916: preferred targets and evidence for conjugative transfer of a single strand and for a double-stranded circular intermediate. Mol Microbiol. 1994 Mar;11(6):1099–1108. doi: 10.1111/j.1365-2958.1994.tb00386.x. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Scott J. R. Sex and the single circle: conjugative transposition. J Bacteriol. 1992 Oct;174(19):6005–6010. doi: 10.1128/jb.174.19.6005-6010.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Su Y. A., Clewell D. B. Characterization of the left 4 kb of conjugative transposon Tn916: determinants involved in excision. Plasmid. 1993 Nov;30(3):234–250. doi: 10.1006/plas.1993.1055. [DOI] [PubMed] [Google Scholar]
  28. Su Y. A., He P., Clewell D. B. Characterization of the tet(M) determinant of Tn916: evidence for regulation by transcription attenuation. Antimicrob Agents Chemother. 1992 Apr;36(4):769–778. doi: 10.1128/aac.36.4.769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Su Y. A., Sulavik M. C., He P., Makinen K. K., Makinen P. L., Fiedler S., Wirth R., Clewell D. B. Nucleotide sequence of the gelatinase gene (gelE) from Enterococcus faecalis subsp. liquefaciens. Infect Immun. 1991 Jan;59(1):415–420. doi: 10.1128/iai.59.1.415-420.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Trieu-Cuot P., Carlier C., Poyart-Salmeron C., Courvalin P. An integrative vector exploiting the transposition properties of Tn1545 for insertional mutagenesis and cloning of genes from gram-positive bacteria. Gene. 1991 Sep 30;106(1):21–27. doi: 10.1016/0378-1119(91)90561-o. [DOI] [PubMed] [Google Scholar]
  31. Wang A., Macrina F. L. Streptococcal plasmid pIP501 has a functional oriT site. J Bacteriol. 1995 Aug;177(15):4199–4206. doi: 10.1128/jb.177.15.4199-4206.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]
  33. Wirth R., An F. Y., Clewell D. B. Highly efficient protoplast transformation system for Streptococcus faecalis and a new Escherichia coli-S. faecalis shuttle vector. J Bacteriol. 1986 Mar;165(3):831–836. doi: 10.1128/jb.165.3.831-836.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yagi Y., Clewell D. B. Recombination-deficient mutant of Streptococcus faecalis. J Bacteriol. 1980 Aug;143(2):966–970. doi: 10.1128/jb.143.2.966-970.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES