Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1990 Mar;172(3):1368–1373. doi: 10.1128/jb.172.3.1368-1373.1990

Integration host factor plays a role in IS50 and Tn5 transposition.

J C Makris 1, P L Nordmann 1, W S Reznikoff 1
PMCID: PMC208607  PMID: 2155200

Abstract

In Escherichia coli, the frequencies of IS50 and Tn5 transposition are greater in Dam- cells than in isogenic Dam+ cells. IS50 transposition is increased approximately 1,000-fold and Tn5 transposition frequencies are increased about 5- to 10-fold in the absence of Dam methylation. However, in cells that are deficient for both integration host factor (IHF) and Dam methylase, the transposition frequencies of IS50 and Tn5 approximate those found in wild-type cells. The absence of IHF alone has no effect on either IS50 or Tn5 transposition. These results suggest that IHF is required for the increased transposition frequencies of IS50 and Tn5 that are observed in Dam- cells. It is also shown that the level of expression of IS50-encoded proteins, P1 and P2, required for IS50 and Tn5 transposition and its regulation does not decrease in IHF- or in IHF- Dam- cells. This result suggests that the effects of IHF on IS50 and Tn5 transposition are not at the level of IS50 gene expression. Finally, IHF is demonstrated to significantly retard the electrophoretic mobility of a 289-base-pair segment of IS50 DNA that contains a putative IHF protein-binding site. The physiological role of this IHF binding site remains to be determined.

Full text

PDF
1368

Images in this article

1371FIG. 5
on p.1371

Selected References

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

  1. Bramhill D., Kornberg A. Duplex opening by dnaA protein at novel sequences in initiation of replication at the origin of the E. coli chromosome. Cell. 1988 Mar 11;52(5):743–755. doi: 10.1016/0092-8674(88)90412-6. [DOI] [PubMed] [Google Scholar]
  2. Dodson K. W., Berg D. E. Factors affecting transposition activity of IS50 and Tn5 ends. Gene. 1989;76(2):207–213. doi: 10.1016/0378-1119(89)90161-3. [DOI] [PubMed] [Google Scholar]
  3. Dodson K. W., Berg D. E. Saturation mutagenesis of the inside end of insertion sequence IS50. Gene. 1989 Dec 21;85(1):75–81. doi: 10.1016/0378-1119(89)90466-6. [DOI] [PubMed] [Google Scholar]
  4. Filutowicz M., Appelt K. The integration host factor of Escherichia coli binds to multiple sites at plasmid R6K gamma origin and is essential for replication. Nucleic Acids Res. 1988 May 11;16(9):3829–3843. doi: 10.1093/nar/16.9.3829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Flamm E. L., Weisberg R. A. Primary structure of the hip gene of Escherichia coli and of its product, the beta subunit of integration host factor. J Mol Biol. 1985 May 25;183(2):117–128. doi: 10.1016/0022-2836(85)90206-2. [DOI] [PubMed] [Google Scholar]
  6. Friden P., Voelkel K., Sternglanz R., Freundlich M. Reduced expression of the isoleucine and valine enzymes in integration host factor mutants of Escherichia coli. J Mol Biol. 1984 Feb 5;172(4):573–579. doi: 10.1016/s0022-2836(84)80024-8. [DOI] [PubMed] [Google Scholar]
  7. Friedman D. I. Integration host factor: a protein for all reasons. Cell. 1988 Nov 18;55(4):545–554. doi: 10.1016/0092-8674(88)90213-9. [DOI] [PubMed] [Google Scholar]
  8. Friedman D. I., Olson E. J., Carver D., Gellert M. Synergistic effect of himA and gyrB mutations: evidence that him functions control expression of ilv and xyl genes. J Bacteriol. 1984 Feb;157(2):484–489. doi: 10.1128/jb.157.2.484-489.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gamas P., Chandler M. G., Prentki P., Galas D. J. Escherichia coli integration host factor binds specifically to the ends of the insertion sequence IS1 and to its major insertion hot-spot in pBR322. J Mol Biol. 1987 May 20;195(2):261–272. doi: 10.1016/0022-2836(87)90648-6. [DOI] [PubMed] [Google Scholar]
  10. Granston A. E., Alessi D. M., Eades L. J., Friedman D. I. A point mutation in the Nul gene of bacteriophage lambda facilitates phage growth in Escherichia coli with himA and gyrB mutations. Mol Gen Genet. 1988 Apr;212(1):149–156. doi: 10.1007/BF00322458. [DOI] [PubMed] [Google Scholar]
  11. Greenstein D., Zinder N. D., Horiuchi K. Integration host factor interacts with the DNA replication enhancer of filamentous phage f1. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6262–6266. doi: 10.1073/pnas.85.17.6262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Isberg R. R., Lazaar A. L., Syvanen M. Regulation of Tn5 by the right-repeat proteins: control at the level of the transposition reaction? Cell. 1982 Oct;30(3):883–892. doi: 10.1016/0092-8674(82)90293-8. [DOI] [PubMed] [Google Scholar]
  13. Isberg R. R., Syvanen M. DNA gyrase is a host factor required for transposition of Tn5. Cell. 1982 Aug;30(1):9–18. doi: 10.1016/0092-8674(82)90006-x. [DOI] [PubMed] [Google Scholar]
  14. Johnson R. C., Reznikoff W. S. Role of the IS50 R proteins in the promotion and control of Tn5 transposition. J Mol Biol. 1984 Aug 25;177(4):645–661. doi: 10.1016/0022-2836(84)90042-1. [DOI] [PubMed] [Google Scholar]
  15. Johnson R. C., Yin J. C., Reznikoff W. S. Control of Tn5 transposition in Escherichia coli is mediated by protein from the right repeat. Cell. 1982 Oct;30(3):873–882. doi: 10.1016/0092-8674(82)90292-6. [DOI] [PubMed] [Google Scholar]
  16. Kennedy M., Chandler M., Lane D. Mapping and regulation of the pifC promoter of the F plasmid. Biochim Biophys Acta. 1988 May 6;950(1):75–80. doi: 10.1016/0167-4781(88)90075-9. [DOI] [PubMed] [Google Scholar]
  17. Krause H. M., Higgins N. P. Positive and negative regulation of the Mu operator by Mu repressor and Escherichia coli integration host factor. J Biol Chem. 1986 Mar 15;261(8):3744–3752. [PubMed] [Google Scholar]
  18. Krebs M. P., Reznikoff W. S. Use of a Tn5 derivative that creates lacZ translational fusions to obtain a transposition mutant. Gene. 1988 Mar 31;63(2):277–285. doi: 10.1016/0378-1119(88)90531-8. [DOI] [PubMed] [Google Scholar]
  19. Kur J., Hasan N., Szybalski W. Physical and biological consequences of interactions between integration host factor (IHF) and coliphage lambda late p'R promoter and its mutants. Gene. 1989 Sep 1;81(1):1–15. doi: 10.1016/0378-1119(89)90331-4. [DOI] [PubMed] [Google Scholar]
  20. Leong J. M., Nunes-Düby S., Lesser C. F., Youderian P., Susskind M. M., Landy A. The phi 80 and P22 attachment sites. Primary structure and interaction with Escherichia coli integration host factor. J Biol Chem. 1985 Apr 10;260(7):4468–4477. [PubMed] [Google Scholar]
  21. Makris J. C., Nordmann P. L., Reznikoff W. S. Mutational analysis of insertion sequence 50 (IS50) and transposon 5 (Tn5) ends. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2224–2228. doi: 10.1073/pnas.85.7.2224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Makris J. C., Reznikoff W. S. Orientation of IS50 transposase gene and IS50 transposition. J Bacteriol. 1989 Sep;171(9):5212–5214. doi: 10.1128/jb.171.9.5212-5214.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mazodier P., Cossart P., Giraud E., Gasser F. Completion of the nucleotide sequence of the central region of Tn5 confirms the presence of three resistance genes. Nucleic Acids Res. 1985 Jan 11;13(1):195–205. doi: 10.1093/nar/13.1.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McCommas S. A., Syvanen M. Temporal control of transposition in Tn5. J Bacteriol. 1988 Feb;170(2):889–894. doi: 10.1128/jb.170.2.889-894.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Miller H. I., Kikuchi A., Nash H. A., Weisberg R. A., Friedman D. I. Site-specific recombination of bacteriophage lambda: the role of host gene products. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1121–1126. doi: 10.1101/sqb.1979.043.01.125. [DOI] [PubMed] [Google Scholar]
  26. Miller H. I. Multilevel regulation of bacteriophage lambda lysogeny by the E. coli himA gene. Cell. 1981 Jul;25(1):269–276. doi: 10.1016/0092-8674(81)90252-x. [DOI] [PubMed] [Google Scholar]
  27. Miller H. I. Primary structure of the himA gene of Escherichia coli: homology with DNA-binding protein HU and association with the phenylalanyl-tRNA synthetase operon. Cold Spring Harb Symp Quant Biol. 1984;49:691–698. doi: 10.1101/sqb.1984.049.01.078. [DOI] [PubMed] [Google Scholar]
  28. Morisato D., Kleckner N. Tn10 transposition and circle formation in vitro. Cell. 1987 Oct 9;51(1):101–111. doi: 10.1016/0092-8674(87)90014-6. [DOI] [PubMed] [Google Scholar]
  29. Pettijohn D. E. Histone-like proteins and bacterial chromosome structure. J Biol Chem. 1988 Sep 15;263(26):12793–12796. [PubMed] [Google Scholar]
  30. Phadnis S. H., Berg D. E. Identification of base pairs in the outside end of insertion sequence IS50 that are needed for IS50 and Tn5 transposition. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9118–9122. doi: 10.1073/pnas.84.24.9118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Russell D. W., Zinder N. D. Hemimethylation prevents DNA replication in E. coli. Cell. 1987 Sep 25;50(7):1071–1079. doi: 10.1016/0092-8674(87)90173-5. [DOI] [PubMed] [Google Scholar]
  32. Sasakawa C., Berg D. E. IS50-mediated inverse transposition. Discrimination between the two ends of an IS element. J Mol Biol. 1982 Aug 5;159(2):257–271. doi: 10.1016/0022-2836(82)90495-8. [DOI] [PubMed] [Google Scholar]
  33. Sasakawa C., Uno Y., Yoshikawa M. The requirement for both DNA polymerase and 5' to 3' exonuclease activities of DNA polymerase I during Tn5 transposition. Mol Gen Genet. 1981;182(1):19–24. doi: 10.1007/BF00422761. [DOI] [PubMed] [Google Scholar]
  34. Sasakawa C., Uno Y., Yoshikawa M. lon-sulA regulatory function affects the efficiency of transposition of Tn5 from lambda b221 cI857 Pam Oam to the chromosome. Biochem Biophys Res Commun. 1987 Feb 13;142(3):879–884. doi: 10.1016/0006-291x(87)91495-1. [DOI] [PubMed] [Google Scholar]
  35. Surette M. G., Lavoie B. D., Chaconas G. Action at a distance in Mu DNA transposition: an enhancer-like element is the site of action of supercoiling relief activity by integration host factor (IHF). EMBO J. 1989 Nov;8(11):3483–3489. doi: 10.1002/j.1460-2075.1989.tb08513.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Syvanen M., Hopkins J. D., Clements M. A new class of mutants in DNA polymerase I that affects gene transposition. J Mol Biol. 1982 Jun 25;158(2):203–212. doi: 10.1016/0022-2836(82)90429-6. [DOI] [PubMed] [Google Scholar]
  37. Yin J. C., Krebs M. P., Reznikoff W. S. Effect of dam methylation on Tn5 transposition. J Mol Biol. 1988 Jan 5;199(1):35–45. doi: 10.1016/0022-2836(88)90377-4. [DOI] [PubMed] [Google Scholar]
  38. Yin J. C., Reznikoff W. S. dnaA, an essential host gene, and Tn5 transposition. J Bacteriol. 1987 Oct;169(10):4637–4645. doi: 10.1128/jb.169.10.4637-4645.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES