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. 1997 Feb;145(2):267–279. doi: 10.1093/genetics/145.2.267

Mobilization of a Minos Transposon in Drosophila Melanogaster Chromosomes and Chromatid Repair by Heteroduplex Formation

B Arca 1, S Zabalou 1, T G Loukeris 1, C Savakis 1
PMCID: PMC1207794  PMID: 9071583

Abstract

Transposase-mediated mobilization of the element Minos has been studied in the Drosophila melanogaster genome. Excision and transposition of a nonautonomous Minos transposon in the presence of a Minos transposase gene was detected with a dominant eye color marker carried by the transposon. Frequencies of excision in somatic tissues and in the germ line were higher in flies heterozygous for the transposon than in homozygotes or hemizygotes. Transposition of a X chromosome-linked insertion of Minos into new autosomal sites occurred in 1-12% of males expressing transposase, suggesting that this system is usable for gene tagging and enhancer trapping in Drosophila. Sequence analysis of PCR-amplified donor sites after excision showed precise restoration of the original target sequence in ~75% of events in heterozygotes and the presence of footprints or partially deleted elements in the remaining events. Most footprints consisted of the four terminal bases of the transposon, flanked by the TA target duplication. Sequencing of a chromosomal donor site that was directly cloned after excision showed a characteristic two-base mismatch heteroduplex in the center of the 6-bp footprint. Circular extrachromosomal forms of the transposon, presumably representing excised Minos elements, could be detected only in the presence of transposase. A model for chromatid repair after Minos excision is discussed in which staggered cuts are first produced at the ends of the inverted repeats, the broken chromatid ends are joined, and the resulting heteroduplex is subsequently repaired. The model also suggests a simple mechanism for the production of the target site duplication and for regeneration of the transposon ends during reintegration.

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Selected References

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  1. Abad P., Quiles C., Tares S., Piotte C., Castagnone-Sereno P., Abadon M., Dalmasso A. Sequences homologous to Tc(s) transposable elements of Caenorhabditis elegans are widely distributed in the phylum nematoda. J Mol Evol. 1991 Sep;33(3):251–258. doi: 10.1007/BF02100676. [DOI] [PubMed] [Google Scholar]
  2. Atkinson P. W., Warren W. D., O'Brochta D. A. The hobo transposable element of Drosophila can be cross-mobilized in houseflies and excises like the Ac element of maize. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9693–9697. doi: 10.1073/pnas.90.20.9693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brezinsky L., Wang G. V., Humphreys T., Hunt J. The transposable element Uhu from Hawaiian Drosophila--member of the widely dispersed class of Tc1-like transposons. Nucleic Acids Res. 1990 Apr 25;18(8):2053–2059. doi: 10.1093/nar/18.8.2053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brierley H. L., Potter S. S. Distinct characteristics of loop sequences of two Drosophila foldback transposable elements. Nucleic Acids Res. 1985 Jan 25;13(2):485–500. doi: 10.1093/nar/13.2.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bryan G., Garza D., Hartl D. Insertion and excision of the transposable element mariner in Drosophila. Genetics. 1990 May;125(1):103–114. doi: 10.1093/genetics/125.1.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Caizzi R., Caggese C., Pimpinelli S. Bari-1, a new transposon-like family in Drosophila melanogaster with a unique heterochromatic organization. Genetics. 1993 Feb;133(2):335–345. doi: 10.1093/genetics/133.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Calvi B. R., Hong T. J., Findley S. D., Gelbart W. M. Evidence for a common evolutionary origin of inverted repeat transposons in Drosophila and plants: hobo, Activator, and Tam3. Cell. 1991 Aug 9;66(3):465–471. doi: 10.1016/0092-8674(81)90010-6. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Coates C. J., Turney C. L., Frommer M., O'Brochta D. A., Warren W. D., Atkinson P. W. The transposable element mariner can excise in non-drosophilid insects. Mol Gen Genet. 1995 Nov 15;249(2):246–252. doi: 10.1007/BF00290372. [DOI] [PubMed] [Google Scholar]
  10. Collins J., Forbes E., Anderson P. The Tc3 family of transposable genetic elements in Caenorhabditis elegans. Genetics. 1989 Jan;121(1):47–55. doi: 10.1093/genetics/121.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Daboussi M. J., Langin T., Brygoo Y. Fot1, a new family of fungal transposable elements. Mol Gen Genet. 1992 Mar;232(1):12–16. doi: 10.1007/BF00299131. [DOI] [PubMed] [Google Scholar]
  12. Doak T. G., Doerder F. P., Jahn C. L., Herrick G. A proposed superfamily of transposase genes: transposon-like elements in ciliated protozoa and a common "D35E" motif. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):942–946. doi: 10.1073/pnas.91.3.942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eide D., Anderson P. Insertion and excision of Caenorhabditis elegans transposable element Tc1. Mol Cell Biol. 1988 Feb;8(2):737–746. doi: 10.1128/mcb.8.2.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Engels W. R., Johnson-Schlitz D. M., Eggleston W. B., Sved J. High-frequency P element loss in Drosophila is homolog dependent. Cell. 1990 Aug 10;62(3):515–525. doi: 10.1016/0092-8674(90)90016-8. [DOI] [PubMed] [Google Scholar]
  15. Engels W. R., Preston C. R., Johnson-Schlitz D. M. Long-range cis preference in DNA homology search over the length of a Drosophila chromosome. Science. 1994 Mar 18;263(5153):1623–1625. doi: 10.1126/science.8128250. [DOI] [PubMed] [Google Scholar]
  16. Finnegan D. J. Eukaryotic transposable elements and genome evolution. Trends Genet. 1989 Apr;5(4):103–107. doi: 10.1016/0168-9525(89)90039-5. [DOI] [PubMed] [Google Scholar]
  17. Franz G., Loukeris T. G., Dialektaki G., Thompson C. R., Savakis C. Mobile Minos elements from Drosophila hydei encode a two-exon transposase with similarity to the paired DNA-binding domain. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4746–4750. doi: 10.1073/pnas.91.11.4746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Franz G., Savakis C. Minos, a new transposable element from Drosophila hydei, is a member of the Tc1-like family of transposons. Nucleic Acids Res. 1991 Dec 11;19(23):6646–6646. doi: 10.1093/nar/19.23.6646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gepner J., Hays T. S. A fertility region on the Y chromosome of Drosophila melanogaster encodes a dynein microtubule motor. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11132–11136. doi: 10.1073/pnas.90.23.11132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hagemann A. T., Craig N. L. Tn7 transposition creates a hotspot for homologous recombination at the transposon donor site. Genetics. 1993 Jan;133(1):9–16. doi: 10.1093/genetics/133.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Harris L. J., Prasad S., Rose A. M. Isolation and sequence analysis of Caenorhabditis briggsae repetitive elements related to the Caenorhabditis elegans transposon Tc1. J Mol Evol. 1990 Apr;30(4):359–369. doi: 10.1007/BF02101890. [DOI] [PubMed] [Google Scholar]
  22. Heierhorst J., Lederis K., Richter D. Presence of a member of the Tc1-like transposon family from nematodes and Drosophila within the vasotocin gene of a primitive vertebrate, the Pacific hagfish Eptatretus stouti. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6798–6802. doi: 10.1073/pnas.89.15.6798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Henikoff S. Detection of Caenorhabditis transposon homologs in diverse organisms. New Biol. 1992 Apr;4(4):382–388. [PubMed] [Google Scholar]
  24. Henikoff S., Henikoff J. G. Amino acid substitution matrices from protein blocks. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10915–10919. doi: 10.1073/pnas.89.22.10915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Izsvák Z., Ivics Z., Hackett P. B. Characterization of a Tc1-like transposable element in zebrafish (Danio rerio). Mol Gen Genet. 1995 May 10;247(3):312–322. doi: 10.1007/BF00293199. [DOI] [PubMed] [Google Scholar]
  26. Jaraczewski J. W., Jahn C. L. Elimination of Tec elements involves a novel excision process. Genes Dev. 1993 Jan;7(1):95–105. doi: 10.1101/gad.7.1.95. [DOI] [PubMed] [Google Scholar]
  27. Jehle J. A., Fritsch E., Nickel A., Huber J., Backhaus H. TCl4.7: a novel lepidopteran transposon found in Cydia pomonella granulosis virus. Virology. 1995 Mar 10;207(2):369–379. doi: 10.1006/viro.1995.1096. [DOI] [PubMed] [Google Scholar]
  28. Kachroo P., Leong S. A., Chattoo B. B. Pot2, an inverted repeat transposon from the rice blast fungus Magnaporthe grisea. Mol Gen Genet. 1994 Nov 1;245(3):339–348. doi: 10.1007/BF00290114. [DOI] [PubMed] [Google Scholar]
  29. Kaufman P. D., Rio D. C. P element transposition in vitro proceeds by a cut-and-paste mechanism and uses GTP as a cofactor. Cell. 1992 Apr 3;69(1):27–39. doi: 10.1016/0092-8674(92)90116-t. [DOI] [PubMed] [Google Scholar]
  30. Kidwell M. G., Kidwell J. F., Sved J. A. Hybrid Dysgenesis in DROSOPHILA MELANOGASTER: A Syndrome of Aberrant Traits Including Mutation, Sterility and Male Recombination. Genetics. 1977 Aug;86(4):813–833. doi: 10.1093/genetics/86.4.813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Koenen M. Recovery of DNA from agarose gels using liquid nitrogen. Trends Genet. 1989 May;5(5):137–137. doi: 10.1016/0168-9525(89)90051-6. [DOI] [PubMed] [Google Scholar]
  32. Kulkosky J., Jones K. S., Katz R. A., Mack J. P., Skalka A. M. Residues critical for retroviral integrative recombination in a region that is highly conserved among retroviral/retrotransposon integrases and bacterial insertion sequence transposases. Mol Cell Biol. 1992 May;12(5):2331–2338. doi: 10.1128/mcb.12.5.2331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lam W. L., Seo P., Robison K., Virk S., Gilbert W. Discovery of amphibian Tc1-like transposon families. J Mol Biol. 1996 Mar 29;257(2):359–366. doi: 10.1006/jmbi.1996.0168. [DOI] [PubMed] [Google Scholar]
  34. Langin T., Capy P., Daboussi M. J. The transposable element impala, a fungal member of the Tc1-mariner superfamily. Mol Gen Genet. 1995 Jan 6;246(1):19–28. doi: 10.1007/BF00290129. [DOI] [PubMed] [Google Scholar]
  35. Levitt A., Emmons S. W. The Tc2 transposon in Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1989 May;86(9):3232–3236. doi: 10.1073/pnas.86.9.3232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Merriman P. J., Grimes C. D., Ambroziak J., Hackett D. A., Skinner P., Simmons M. J. S elements: a family of Tc1-like transposons in the genome of Drosophila melanogaster. Genetics. 1995 Dec;141(4):1425–1438. doi: 10.1093/genetics/141.4.1425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Picard G., Bregliano J. C., Bucheton A., Lavige J. M., Pelisson A., Kidwell M. G. Non-mendelian female sterility and hybrid dysgenesis in Drosophila melanogaster. Genet Res. 1978 Nov;32(3):275–287. doi: 10.1017/s0016672300018772. [DOI] [PubMed] [Google Scholar]
  39. Pirrotta V. Vectors for P-mediated transformation in Drosophila. Biotechnology. 1988;10:437–456. doi: 10.1016/b978-0-409-90042-2.50028-3. [DOI] [PubMed] [Google Scholar]
  40. Plasterk R. H. The origin of footprints of the Tc1 transposon of Caenorhabditis elegans. EMBO J. 1991 Jul;10(7):1919–1925. doi: 10.1002/j.1460-2075.1991.tb07718.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Polard P., Prère M. F., Fayet O., Chandler M. Transposase-induced excision and circularization of the bacterial insertion sequence IS911. EMBO J. 1992 Dec;11(13):5079–5090. doi: 10.1002/j.1460-2075.1992.tb05615.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Radice A. D., Emmons S. W. Extrachromosomal circular copies of the transposon Tc1. Nucleic Acids Res. 1993 Jun 11;21(11):2663–2667. doi: 10.1093/nar/21.11.2663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Robertson H. M., Lampe D. J. Distribution of transposable elements in arthropods. Annu Rev Entomol. 1995;40:333–357. doi: 10.1146/annurev.en.40.010195.002001. [DOI] [PubMed] [Google Scholar]
  44. Robertson H. M., MacLeod E. G. Five major subfamilies of mariner transposable elements in insects, including the Mediterranean fruit fly, and related arthropods. Insect Mol Biol. 1993;2(3):125–139. doi: 10.1111/j.1365-2583.1993.tb00132.x. [DOI] [PubMed] [Google Scholar]
  45. Rose A. M., Snutch T. P. Isolation of the closed circular form of the transposable element Tc1 in Caenorhabditis elegans. Nature. 1984 Oct 4;311(5985):485–486. doi: 10.1038/311485a0. [DOI] [PubMed] [Google Scholar]
  46. Ruan K. S., Emmons S. W. Precise and imprecise somatic excision of the transposon Tc1 in the nematode C. elegans. Nucleic Acids Res. 1987 Sep 11;15(17):6875–6881. doi: 10.1093/nar/15.17.6875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Ruan K., Emmons S. W. Extrachromosomal copies of transposon Tc1 in the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4018–4022. doi: 10.1073/pnas.81.13.4018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Rubin G. M., Spradling A. C. Genetic transformation of Drosophila with transposable element vectors. Science. 1982 Oct 22;218(4570):348–353. doi: 10.1126/science.6289436. [DOI] [PubMed] [Google Scholar]
  49. Saedler H., Nevers P. Transposition in plants: a molecular model. EMBO J. 1985 Mar;4(3):585–590. doi: 10.1002/j.1460-2075.1985.tb03670.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  51. Staveley B. E., Heslip T. R., Hodgetts R. B., Bell J. B. Protected P-element termini suggest a role for inverted-repeat-binding protein in transposase-induced gap repair in Drosophila melanogaster. Genetics. 1995 Mar;139(3):1321–1329. doi: 10.1093/genetics/139.3.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Takasu-Ishikawa E., Yoshihara M., Hotta Y. Extra sequences found at P element excision sites in Drosophila melanogaster. Mol Gen Genet. 1992 Mar;232(1):17–23. doi: 10.1007/BF00299132. [DOI] [PubMed] [Google Scholar]
  53. Turlan C., Chandler M. IS1-mediated intramolecular rearrangements: formation of excised transposon circles and replicative deletions. EMBO J. 1995 Nov 1;14(21):5410–5421. doi: 10.1002/j.1460-2075.1995.tb00225.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Vos J. C., De Baere I., Plasterk R. H. Transposase is the only nematode protein required for in vitro transposition of Tc1. Genes Dev. 1996 Mar 15;10(6):755–761. doi: 10.1101/gad.10.6.755. [DOI] [PubMed] [Google Scholar]
  55. Williams K., Doak T. G., Herrick G. Developmental precise excision of Oxytricha trifallax telomere-bearing elements and formation of circles closed by a copy of the flanking target duplication. EMBO J. 1993 Dec;12(12):4593–4601. doi: 10.1002/j.1460-2075.1993.tb06148.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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