Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 Feb;170(2):889–894. doi: 10.1128/jb.170.2.889-894.1988

Temporal control of transposition in Tn5.

S A McCommas 1, M Syvanen 1
PMCID: PMC210738  PMID: 2828332

Abstract

IS50R is an insertion sequence associated with the transposon Tn5. IS50R carries the structural genes for two proteins; one (P1) is the Tn5 transposase, and the other (P2) is an inhibitor of transposition. These two proteins are translated from two different transcripts, m1 and m2. When bacteriophage lambda::IS50R DNA was introduced into a bacterial cell, m1 and m2 were initially at relative levels of about 1 to 2. As time progressed the amount of m1 fell, whereas the amount of m2 continued to increase, until after about 3 h the ratio of m1 to m2 was about 1 to 80. The temporal changes in the levels of these transcripts correlated with temporal changes in P1 and P2 levels and Tn5 transposition that have been documented in other studies. We measured the stability of the messages and showed that the differences in the levels of m1 and m2 must reflect real differences in the strengths of their promoters and that the changes in transcription kinetics are mediated by the dam methylation system of the cell and are not determined by IS50R products. Our results show that the 5' end of m2 is about twice as stable as that of m1, which raises the possibility that differential message stability does, in part, influence the ratio of inhibitor to transposase.

Full text

PDF
889

Images in this article

891Image
on p.891
892Image
on p.892

Selected References

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

  1. Auerswald E. A., Ludwig G., Schaller H. Structural analysis of Tn5. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):107–113. doi: 10.1101/sqb.1981.045.01.019. [DOI] [PubMed] [Google Scholar]
  2. Barbeyron T., Kean K., Forterre P. DNA adenine methylation of GATC sequences appeared recently in the Escherichia coli lineage. J Bacteriol. 1984 Nov;160(2):586–590. doi: 10.1128/jb.160.2.586-590.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Belasco J. G., Nilsson G., von Gabain A., Cohen S. N. The stability of E. coli gene transcripts is dependent on determinants localized to specific mRNA segments. Cell. 1986 Jul 18;46(2):245–251. doi: 10.1016/0092-8674(86)90741-5. [DOI] [PubMed] [Google Scholar]
  4. Berg D. E., Johnsrud L., McDivitt L., Ramabhadran R., Hirschel B. J. Inverted repeats of Tn5 are transposable elements. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2632–2635. doi: 10.1073/pnas.79.8.2632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  6. Biek D., Roth J. R. Regulation of Tn5 transposition in Salmonella typhimurium. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6047–6051. doi: 10.1073/pnas.77.10.6047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Duesberg P. H., Vogt P. K. Gel electrophoresis of avian leukosis and sarcoma viral RNA in formamide: comparison with other viral and cellular RNA species. J Virol. 1973 Sep;12(3):594–599. doi: 10.1128/jvi.12.3.594-599.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Isberg R. R., Syvanen M. Compartmentalization of the proteins encoded by IS50R. J Biol Chem. 1985 Mar 25;260(6):3645–3651. [PubMed] [Google Scholar]
  10. 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]
  11. Isberg R. R., Syvanen M. Replicon fusions promoted by the inverted repeats of Tn5. The right repeat is an insertion sequence. J Mol Biol. 1981 Jul 25;150(1):15–32. doi: 10.1016/0022-2836(81)90322-3. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Krebs M. P., Reznikoff W. S. Transcriptional and translational initiation sites of IS50. Control of transposase and inhibitor expression. J Mol Biol. 1986 Dec 20;192(4):781–791. doi: 10.1016/0022-2836(86)90028-8. [DOI] [PubMed] [Google Scholar]
  15. Lyons S. M., Schendel P. F. Kinetics of methylation in Escherichia coli K-12. J Bacteriol. 1984 Jul;159(1):421–423. doi: 10.1128/jb.159.1.421-423.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mazodier P., Genilloud O., Giraud E., Gasser F. Expression of Tn5-encoded streptomycin resistance in E. coli. Mol Gen Genet. 1986 Sep;204(3):404–409. doi: 10.1007/BF00331016. [DOI] [PubMed] [Google Scholar]
  17. Nilsson G., Belasco J. G., Cohen S. N., von Gabain A. Growth-rate dependent regulation of mRNA stability in Escherichia coli. Nature. 1984 Nov 1;312(5989):75–77. doi: 10.1038/312075a0. [DOI] [PubMed] [Google Scholar]
  18. Roberts D., Hoopes B. C., McClure W. R., Kleckner N. IS10 transposition is regulated by DNA adenine methylation. Cell. 1985 Nov;43(1):117–130. doi: 10.1016/0092-8674(85)90017-0. [DOI] [PubMed] [Google Scholar]
  19. Rossetti O. L., Altman R., Young R. Kinetics of Tn5 transposition. Gene. 1984 Dec;32(1-2):91–98. doi: 10.1016/0378-1119(84)90036-2. [DOI] [PubMed] [Google Scholar]
  20. Rothstein S. J., Reznikoff W. S. The functional differences in the inverted repeats of Tn5 are caused by a single base pair nonhomology. Cell. 1981 Jan;23(1):191–199. doi: 10.1016/0092-8674(81)90284-1. [DOI] [PubMed] [Google Scholar]
  21. SCHERRER K., DARNELL J. E. Sedimentation characteristics of rapidly labelled RNA from HeLa cells. Biochem Biophys Res Commun. 1962 Jun 4;7:486–490. doi: 10.1016/0006-291x(62)90341-8. [DOI] [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. Syvanen M. The evolutionary implications of mobile genetic elements. Annu Rev Genet. 1984;18:271–293. doi: 10.1146/annurev.ge.18.120184.001415. [DOI] [PubMed] [Google Scholar]
  24. de Boer H. A., Comstock L. J., Vasser M. The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci U S A. 1983 Jan;80(1):21–25. doi: 10.1073/pnas.80.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. von Gabain A., Belasco J. G., Schottel J. L., Chang A. C., Cohen S. N. Decay of mRNA in Escherichia coli: investigation of the fate of specific segments of transcripts. Proc Natl Acad Sci U S A. 1983 Feb;80(3):653–657. doi: 10.1073/pnas.80.3.653. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES