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. 1997 Apr;179(8):2567–2572. doi: 10.1128/jb.179.8.2567-2572.1997

DNA binding by the Xis protein of the conjugative transposon Tn916.

C K Rudy 1, J R Scott 1, G Churchward 1
PMCID: PMC179005  PMID: 9098054

Abstract

We purified the Xis protein of the conjugative transposon Tn916 and showed by nuclease protection experiments that Xis bound specifically to sites close to each end of Tn916. These specific binding sites are close to, and in the same relative orientation to, binding sites for the N-terminal domain of Tn916 integrase protein. These results suggest that Xis is involved in the formation of nucleoprotein structures at the ends of Tn916 that help to correctly align the ends so that excision can occur.

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Selected References

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  1. Abremski K., Gottesman S. Site-specific recombination Xis-independent excisive recombination of bacteriophage lambda. J Mol Biol. 1981 Nov 25;153(1):67–78. doi: 10.1016/0022-2836(81)90527-1. [DOI] [PubMed] [Google Scholar]
  2. Argos P., Landy A., Abremski K., Egan J. B., Haggard-Ljungquist E., Hoess R. H., Kahn M. L., Kalionis B., Narayana S. V., Pierson L. S., 3rd The integrase family of site-specific recombinases: regional similarities and global diversity. EMBO J. 1986 Feb;5(2):433–440. doi: 10.1002/j.1460-2075.1986.tb04229.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bringel F., Van Alstine G. L., Scott J. R. Conjugative transposition of Tn916: the transposon int gene is required only in the donor. J Bacteriol. 1992 Jun;174(12):4036–4041. doi: 10.1128/jb.174.12.4036-4041.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bushman W., Yin S., Thio L. L., Landy A. Determinants of directionality in lambda site-specific recombination. Cell. 1984 Dec;39(3 Pt 2):699–706. doi: 10.1016/0092-8674(84)90477-x. [DOI] [PubMed] [Google Scholar]
  5. Buu-Hoï A., Horodniceanu T. Conjugative transfer of multiple antibiotic resistance markers in Streptococcus pneumoniae. J Bacteriol. 1980 Jul;143(1):313–320. doi: 10.1128/jb.143.1.313-320.1980. [DOI] [PMC free article] [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. 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]
  8. 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]
  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., 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]
  11. 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]
  12. Franz B., Landy A. The Holliday junction intermediates of lambda integrative and excisive recombination respond differently to the bending proteins integration host factor and excisionase. EMBO J. 1995 Jan 16;14(2):397–406. doi: 10.1002/j.1460-2075.1995.tb07014.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gottesman S. Minimizing proteolysis in Escherichia coli: genetic solutions. Methods Enzymol. 1990;185:119–129. doi: 10.1016/0076-6879(90)85013-e. [DOI] [PubMed] [Google Scholar]
  14. Ike Y., Flannagan S. E., Clewell D. B. Hyperhemolytic phenomena associated with insertions of Tn916 into the hemolysin determinant of Enterococcus faecalis plasmid pAD1. J Bacteriol. 1992 Mar;174(6):1801–1809. doi: 10.1128/jb.174.6.1801-1809.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kim S., Landy A. Lambda Int protein bridges between higher order complexes at two distant chromosomal loci attL and attR. Science. 1992 Apr 10;256(5054):198–203. doi: 10.1126/science.1533056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kim S., Moitoso de Vargas L., Nunes-Düby S. E., Landy A. Mapping of a higher order protein-DNA complex: two kinds of long-range interactions in lambda attL. Cell. 1990 Nov 16;63(4):773–781. doi: 10.1016/0092-8674(90)90143-3. [DOI] [PubMed] [Google Scholar]
  17. Landy A. Dynamic, structural, and regulatory aspects of lambda site-specific recombination. Annu Rev Biochem. 1989;58:913–949. doi: 10.1146/annurev.bi.58.070189.004405. [DOI] [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. Manganelli R., Ricci S., Pozzi G. Conjugative transposon Tn916: evidence for excision with formation of 5'-protruding termini. J Bacteriol. 1996 Oct;178(19):5813–5816. doi: 10.1128/jb.178.19.5813-5816.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moitoso de Vargas L., Landy A. A switch in the formation of alternative DNA loops modulates lambda site-specific recombination. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):588–592. doi: 10.1073/pnas.88.2.588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Numrych T. E., Gumport R. I., Gardner J. F. Characterization of the bacteriophage lambda excisionase (Xis) protein: the C-terminus is required for Xis-integrase cooperativity but not for DNA binding. EMBO J. 1992 Oct;11(10):3797–3806. doi: 10.1002/j.1460-2075.1992.tb05465.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Rauch P. J., De Vos W. M. Characterization of the novel nisin-sucrose conjugative transposon Tn5276 and its insertion in Lactococcus lactis. J Bacteriol. 1992 Feb;174(4):1280–1287. doi: 10.1128/jb.174.4.1280-1287.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rauch P. J., de Vos W. M. Identification and characterization of genes involved in excision of the Lactococcus lactis conjugative transposon Tn5276. J Bacteriol. 1994 Apr;176(8):2165–2171. doi: 10.1128/jb.176.8.2165-2171.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rudy C. K., Scott J. R. Length of the coupling sequence of Tn916. J Bacteriol. 1994 Jun;176(11):3386–3388. doi: 10.1128/jb.176.11.3386-3388.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Scott J. R., Churchward G. G. Conjugative transposition. Annu Rev Microbiol. 1995;49:367–397. doi: 10.1146/annurev.mi.49.100195.002055. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. 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]
  32. 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]
  33. Taylor K. L., Churchward G. Specific DNA cleavage mediated by the integrase of conjugative transposon Tn916. J Bacteriol. 1997 Feb;179(4):1117–1125. doi: 10.1128/jb.179.4.1117-1125.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thompson J. F., Landy A. Empirical estimation of protein-induced DNA bending angles: applications to lambda site-specific recombination complexes. Nucleic Acids Res. 1988 Oct 25;16(20):9687–9705. doi: 10.1093/nar/16.20.9687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yin S., Bushman W., Landy A. Interaction of the lambda site-specific recombination protein Xis with attachment site DNA. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1040–1044. doi: 10.1073/pnas.82.4.1040. [DOI] [PMC free article] [PubMed] [Google Scholar]

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