Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 1998 May;149(1):179–187. doi: 10.1093/genetics/149.1.179

Factors affecting transposition of the Himar1 mariner transposon in vitro.

D J Lampe 1, T E Grant 1, H M Robertson 1
PMCID: PMC1460121  PMID: 9584095

Abstract

Mariner family transposable elements are widespread in animals, but their regulation is poorly understood, partly because only two are known to be functional. These are particular copies of the Dmmar1 element from Drosophila mauritiana, for example, Mos1, and the consensus sequence of the Himar1 element from the horn fly, Haematobia irritans. An in vitro transposition system was refined to investigate several parameters that influence the transposition of Himar1. Transposition products accumulated linearly over a period of 6 hr. Transposition frequency increased with temperature and was dependent on Mg2+ concentration. Transposition frequency peaked over a narrow range of transposase concentration. The decline at higher concentrations, a phenomenon observed in vivo with Mos1, supports the suggestion that mariners may be regulated in part by "overproduction inhibition." Transposition frequency decreased exponentially with increasing transposon size and was affected by the sequence of the flanking DNA of the donor site. A noticeable bias in target site usage suggests a preference for insertion into bent or bendable DNA sequences rather than any specific nucleotide sequences beyond the TA target site.

Full Text

The Full Text of this article is available as a PDF (198.1 KB).

Selected References

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

  1. Ali S. A., Steinkasserer A. PCR-ligation-PCR mutagenesis: a protocol for creating gene fusions and mutations. Biotechniques. 1995 May;18(5):746–750. [PubMed] [Google Scholar]
  2. Arciszewska L. K., Craig N. L. Interaction of the Tn7-encoded transposition protein TnsB with the ends of the transposon. Nucleic Acids Res. 1991 Sep 25;19(18):5021–5029. doi: 10.1093/nar/19.18.5021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benedict M. Q., Salazar C. E., Collins F. H. A new dominant selectable marker for genetic transformation; Hsp70-opd. Insect Biochem Mol Biol. 1995 Dec;25(10):1061–1065. doi: 10.1016/0965-1748(95)00061-5. [DOI] [PubMed] [Google Scholar]
  4. Bigot Y., Hamelin M. H., Capy P., Periquet G. Mariner-like elements in hymenopteran species: insertion site and distribution. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3408–3412. doi: 10.1073/pnas.91.8.3408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brukner I., Sánchez R., Suck D., Pongor S. Sequence-dependent bending propensity of DNA as revealed by DNase I: parameters for trinucleotides. EMBO J. 1995 Apr 18;14(8):1812–1818. doi: 10.1002/j.1460-2075.1995.tb07169.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chalmers R. M., Kleckner N. Tn10/IS10 transposase purification, activation, and in vitro reaction. J Biol Chem. 1994 Mar 18;269(11):8029–8035. [PubMed] [Google Scholar]
  7. Derbyshire K. M., Grindley N. D. Binding of the IS903 transposase to its inverted repeat in vitro. EMBO J. 1992 Sep;11(9):3449–3455. doi: 10.1002/j.1460-2075.1992.tb05424.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Garcia-Fernàndez J., Bayascas-Ramírez J. R., Marfany G., Muñoz-Mármol A. M., Casali A., Baguñ J., Saló E. High copy number of highly similar mariner-like transposons in planarian (Platyhelminthe): evidence for a trans-phyla horizontal transfer. Mol Biol Evol. 1995 May;12(3):421–431. doi: 10.1093/oxfordjournals.molbev.a040217. [DOI] [PubMed] [Google Scholar]
  9. Garza D., Medhora M., Koga A., Hartl D. L. Introduction of the transposable element mariner into the germline of Drosophila melanogaster. Genetics. 1991 Jun;128(2):303–310. doi: 10.1093/genetics/128.2.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gill S. C., von Hippel P. H. Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989 Nov 1;182(2):319–326. doi: 10.1016/0003-2697(89)90602-7. [DOI] [PubMed] [Google Scholar]
  11. Gueiros-Filho F. J., Beverley S. M. Trans-kingdom transposition of the Drosophila element mariner within the protozoan Leishmania. Science. 1997 Jun 13;276(5319):1716–1719. doi: 10.1126/science.276.5319.1716. [DOI] [PubMed] [Google Scholar]
  12. Hagerman P. J. Sequence-directed curvature of DNA. Nature. 1986 May 22;321(6068):449–450. doi: 10.1038/321449a0. [DOI] [PubMed] [Google Scholar]
  13. Hartl D. L., Lozovskaya E. R., Nurminsky D. I., Lohe A. R. What restricts the activity of mariner-like transposable elements. Trends Genet. 1997 May;13(5):197–201. doi: 10.1016/s0168-9525(97)01087-1. [DOI] [PubMed] [Google Scholar]
  14. Keeler K. J., Dray T., Penney J. E., Gloor G. B. Gene targeting of a plasmid-borne sequence to a double-strand DNA break in Drosophila melanogaster. Mol Cell Biol. 1996 Feb;16(2):522–528. doi: 10.1128/mcb.16.2.522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kunze R., Behrens U., Courage-Franzkowiak U., Feldmar S., Kühn S., Lütticke R. Dominant transposition-deficient mutants of maize Activator (Ac) transposase. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7094–7098. doi: 10.1073/pnas.90.15.7094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lampe D. J., Churchill M. E., Robertson H. M. A purified mariner transposase is sufficient to mediate transposition in vitro. EMBO J. 1996 Oct 1;15(19):5470–5479. [PMC free article] [PubMed] [Google Scholar]
  17. Lidholm D. A., Lohe A. R., Hartl D. L. The transposable element mariner mediates germline transformation in Drosophila melanogaster. Genetics. 1993 Jul;134(3):859–868. doi: 10.1093/genetics/134.3.859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lohe A. R., Hartl D. L. Autoregulation of mariner transposase activity by overproduction and dominant-negative complementation. Mol Biol Evol. 1996 Apr;13(4):549–555. doi: 10.1093/oxfordjournals.molbev.a025615. [DOI] [PubMed] [Google Scholar]
  19. Lohe A. R., Hartl D. L. Germline transformation of Drosophila virilis with the transposable element mariner. Genetics. 1996 May;143(1):365–374. doi: 10.1093/genetics/143.1.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lohe A. R., Hartl D. L. Reduced germline mobility of a mariner vector containing exogenous DNA: effect of size or site? Genetics. 1996 Jul;143(3):1299–1306. doi: 10.1093/genetics/143.3.1299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Medhora M. M., MacPeek A. H., Hartl D. L. Excision of the Drosophila transposable element mariner: identification and characterization of the Mos factor. EMBO J. 1988 Jul;7(7):2185–2189. doi: 10.1002/j.1460-2075.1988.tb03057.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Milot E., Belmaaza A., Rassart E., Chartrand P. Association of a host DNA structure with retroviral integration sites in chromosomal DNA. Virology. 1994 Jun;201(2):408–412. doi: 10.1006/viro.1994.1310. [DOI] [PubMed] [Google Scholar]
  23. Müller H. P., Varmus H. E. DNA bending creates favored sites for retroviral integration: an explanation for preferred insertion sites in nucleosomes. EMBO J. 1994 Oct 3;13(19):4704–4714. doi: 10.1002/j.1460-2075.1994.tb06794.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Patton J. S., Gomes X. V., Geyer P. K. Position-independent germline transformation in Drosophila using a cuticle pigmentation gene as a selectable marker. Nucleic Acids Res. 1992 Nov 11;20(21):5859–5860. doi: 10.1093/nar/20.21.5859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Robertson H. M., Asplund M. L. Bmmar1: a basal lineage of the mariner family of transposable elements in the silkworm moth, Bombyx mori. Insect Biochem Mol Biol. 1996 Sep-Oct;26(8-9):945–954. doi: 10.1016/s0965-1748(96)00061-6. [DOI] [PubMed] [Google Scholar]
  26. Robertson H. M., Lampe D. J. Recent horizontal transfer of a mariner transposable element among and between Diptera and Neuroptera. Mol Biol Evol. 1995 Sep;12(5):850–862. doi: 10.1093/oxfordjournals.molbev.a040262. [DOI] [PubMed] [Google Scholar]
  27. Robertson H. M. The mariner transposable element is widespread in insects. Nature. 1993 Mar 18;362(6417):241–245. doi: 10.1038/362241a0. [DOI] [PubMed] [Google Scholar]
  28. Sarkar A., Yardley K., Atkinson P. W., James A. A., O'Brochta D. A. Transposition of the Hermes element in embryos of the vector mosquito, Aedes aegypti. Insect Biochem Mol Biol. 1997 May;27(5):359–363. doi: 10.1016/s0965-1748(97)00018-0. [DOI] [PubMed] [Google Scholar]
  29. Schneider T. D., Stephens R. M. Sequence logos: a new way to display consensus sequences. Nucleic Acids Res. 1990 Oct 25;18(20):6097–6100. doi: 10.1093/nar/18.20.6097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Way J. C., Kleckner N. Transposition of plasmid-borne Tn10 elements does not exhibit simple length-dependence. Genetics. 1985 Dec;111(4):705–713. doi: 10.1093/genetics/111.4.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weinreich M. D., Gasch A., Reznikoff W. S. Evidence that the cis preference of the Tn5 transposase is caused by nonproductive multimerization. Genes Dev. 1994 Oct 1;8(19):2363–2374. doi: 10.1101/gad.8.19.2363. [DOI] [PubMed] [Google Scholar]
  32. Wiegand T. W., Reznikoff W. S. Characterization of two hypertransposing Tn5 mutants. J Bacteriol. 1992 Feb;174(4):1229–1239. doi: 10.1128/jb.174.4.1229-1239.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. York D., Reznikoff W. S. DNA binding and phasing analyses of Tn5 transposase and a monomeric variant. Nucleic Acids Res. 1997 Jun 1;25(11):2153–2160. doi: 10.1093/nar/25.11.2153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. van Pouderoyen G., Ketting R. F., Perrakis A., Plasterk R. H., Sixma T. K. Crystal structure of the specific DNA-binding domain of Tc3 transposase of C.elegans in complex with transposon DNA. EMBO J. 1997 Oct 1;16(19):6044–6054. doi: 10.1093/emboj/16.19.6044. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES