Skip to main content
Genetics logoLink to Genetics
. 1994 Mar;136(3):1001–1011. doi: 10.1093/genetics/136.3.1001

P Element-Mediated in Vivo Deletion Analysis of White-Apricot: Deletions between Direct Repeats Are Strongly Favored

M Kurkulos 1, J M Weinberg 1, D Roy 1, S M Mount 1
PMCID: PMC1205858  PMID: 8005410

Abstract

We have isolated and characterized deletions arising within a P transposon, P[hsw(a)], in the presence of P transposase. P[hsw(a)] carries white-apricot (w(a)) sequences, including a complete copia element, under the control of an hsp70 promoter, and resembles the original w(a) allele in eye color phenotype. In the presence of P transposase, P[hsw(a)] shows a high overall rate (approximately 3%) of germline mutations that result in increased eye pigmentation. Of 234 derivatives of P[hsw(a)] with greatly increased eye pigmentation, at least 205 carried deletions within copia. Of these, 201 were precise deletions between the directly repeated 276-nucleotide copia long terminal repeats (LTRs), and four were unique deletions. High rates of transposase-induced precise deletion were observed within another P transposon carrying unrelated 599 nucleotide repeats (yeast 2μ FLP; recombinase target sites) separated by 5.7 kb. Our observation that P element-mediated deletion formation occurs preferentially between direct repeats suggests general methods for controlling deletion formation.

Full Text

The Full Text of this article is available as a PDF (8.1 MB).

Selected References

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

  1. Allard M. W., Ellsworth D. L., Honeycutt R. L. The production of single-stranded DNA suitable for sequencing using the polymerase chain reaction. Biotechniques. 1991 Jan;10(1):24–26. [PubMed] [Google Scholar]
  2. Ballinger D. G., Benzer S. Targeted gene mutations in Drosophila. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9402–9406. doi: 10.1073/pnas.86.23.9402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Banga S. S., Boyd J. B. Oligonucleotide-directed site-specific mutagenesis in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1735–1739. doi: 10.1073/pnas.89.5.1735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carbonare B. D., Gehring W. J. Excision of copia element in a revertant of the white-apricot mutation of Drosophila melanogaster leaves behind one long-terminal repeat. Mol Gen Genet. 1985;199(1):1–6. doi: 10.1007/BF00327501. [DOI] [PubMed] [Google Scholar]
  5. Daniels S. B., Chovnick A. P element transposition in Drosophila melanogaster: an analysis of sister-chromatid pairs and the formation of intragenic secondary insertions during meiosis. Genetics. 1993 Mar;133(3):623–636. doi: 10.1093/genetics/133.3.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davison D., Chapman C. H., Wedeen C., Bingham P. M. Genetic and physical studies of a portion of the white locus participating in transcriptional regulation and in synapsis-dependent interactions in Drosophila adult tissues. Genetics. 1985 Jul;110(3):479–494. doi: 10.1093/genetics/110.3.479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dreesen T. D., Henikoff S., Loughney K. A pairing-sensitive element that mediates trans-inactivation is associated with the Drosophila brown gene. Genes Dev. 1991 Mar;5(3):331–340. doi: 10.1101/gad.5.3.331. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Formosa T., Alberts B. M. DNA synthesis dependent on genetic recombination: characterization of a reaction catalyzed by purified bacteriophage T4 proteins. Cell. 1986 Dec 5;47(5):793–806. doi: 10.1016/0092-8674(86)90522-2. [DOI] [PubMed] [Google Scholar]
  10. Geyer P. K., Green M. M., Corces V. G. Tissue-specific transcriptional enhancers may act in trans on the gene located in the homologous chromosome: the molecular basis of transvection in Drosophila. EMBO J. 1990 Jul;9(7):2247–2256. doi: 10.1002/j.1460-2075.1990.tb07395.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gloor G. B., Nassif N. A., Johnson-Schlitz D. M., Preston C. R., Engels W. R. Targeted gene replacement in Drosophila via P element-induced gap repair. Science. 1991 Sep 6;253(5024):1110–1117. doi: 10.1126/science.1653452. [DOI] [PubMed] [Google Scholar]
  12. Golic K. G., Lindquist S. The FLP recombinase of yeast catalyzes site-specific recombination in the Drosophila genome. Cell. 1989 Nov 3;59(3):499–509. doi: 10.1016/0092-8674(89)90033-0. [DOI] [PubMed] [Google Scholar]
  13. Golic K. G. Site-specific recombination between homologous chromosomes in Drosophila. Science. 1991 May 17;252(5008):958–961. doi: 10.1126/science.2035025. [DOI] [PubMed] [Google Scholar]
  14. Haber J. E. Exploring the pathways of homologous recombination. Curr Opin Cell Biol. 1992 Jun;4(3):401–412. doi: 10.1016/0955-0674(92)90005-w. [DOI] [PubMed] [Google Scholar]
  15. Hsiao K. A fast and simple procedure for sequencing double stranded DNA with sequenase. Nucleic Acids Res. 1991 May 25;19(10):2787–2787. doi: 10.1093/nar/19.10.2787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Johnson-Schlitz D. M., Engels W. R. P-element-induced interallelic gene conversion of insertions and deletions in Drosophila melanogaster. Mol Cell Biol. 1993 Nov;13(11):7006–7018. doi: 10.1128/mcb.13.11.7006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Karess R. E., Rubin G. M. Analysis of P transposable element functions in Drosophila. Cell. 1984 Aug;38(1):135–146. doi: 10.1016/0092-8674(84)90534-8. [DOI] [PubMed] [Google Scholar]
  18. Kurkulos M., Weinberg J. M., Pepling M. E., Mount S. M. Polyadenylylation in copia requires unusually distant upstream sequences. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3038–3042. doi: 10.1073/pnas.88.8.3038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Laurie-Ahlberg C. C., Stam L. F. Use of P-element-mediated transformation to identify the molecular basis of naturally occurring variants affecting Adh expression in Drosophila melanogaster. Genetics. 1987 Jan;115(1):129–140. doi: 10.1093/genetics/115.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lin F. L., Sperle K., Sternberg N. Intermolecular recombination between DNAs introduced into mouse L cells is mediated by a nonconservative pathway that leads to crossover products. Mol Cell Biol. 1990 Jan;10(1):103–112. doi: 10.1128/mcb.10.1.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mount S. M., Green M. M., Rubin G. M. Partial revertants of the transposable element-associated suppressible allele white-apricot in Drosophila melanogaster: structures and responsiveness to genetic modifiers. Genetics. 1988 Feb;118(2):221–234. doi: 10.1093/genetics/118.2.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mount S. M., Rubin G. M. Complete nucleotide sequence of the Drosophila transposable element copia: homology between copia and retroviral proteins. Mol Cell Biol. 1985 Jul;5(7):1630–1638. doi: 10.1128/mcb.5.7.1630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nassif N., Engels W. DNA homology requirements for mitotic gap repair in Drosophila. Proc Natl Acad Sci U S A. 1993 Feb 15;90(4):1262–1266. doi: 10.1073/pnas.90.4.1262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. O'Hare K., Rubin G. M. Structures of P transposable elements and their sites of insertion and excision in the Drosophila melanogaster genome. Cell. 1983 Aug;34(1):25–35. doi: 10.1016/0092-8674(83)90133-2. [DOI] [PubMed] [Google Scholar]
  25. Robertson H. M., Preston C. R., Phillis R. W., Johnson-Schlitz D. M., Benz W. K., Engels W. R. A stable genomic source of P element transposase in Drosophila melanogaster. Genetics. 1988 Mar;118(3):461–470. doi: 10.1093/genetics/118.3.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Spradling A. C., Rubin G. M. The effect of chromosomal position on the expression of the Drosophila xanthine dehydrogenase gene. Cell. 1983 Aug;34(1):47–57. doi: 10.1016/0092-8674(83)90135-6. [DOI] [PubMed] [Google Scholar]
  28. Steller H., Pirrotta V. Expression of the Drosophila white gene under the control of the hsp70 heat shock promoter. EMBO J. 1985 Dec 30;4(13B):3765–3772. doi: 10.1002/j.1460-2075.1985.tb04146.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Struhl G., Basler K. Organizing activity of wingless protein in Drosophila. Cell. 1993 Feb 26;72(4):527–540. doi: 10.1016/0092-8674(93)90072-x. [DOI] [PubMed] [Google Scholar]
  30. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Xu T., Rubin G. M. Analysis of genetic mosaics in developing and adult Drosophila tissues. Development. 1993 Apr;117(4):1223–1237. doi: 10.1242/dev.117.4.1223. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES