Abstract
We have studied P transposase-induced events on a P[w] transgene, P[w(d1)], harboring the whole white gene with a 3.44-kb direct duplication of its 5' regulatory sequences (containing the ZESTE-binding region, ZBR). We have recovered mutations leading to an increase or a decrease of zeste(1) repression, generally as the consequence of modifications of the number of ZBR in close physical proximity and/or jumps to other sites. We describe mutants displaying deletions of the original duplicated sequence or increases in the number of repeats from two to three or four. Internal deletions are more frequent than amplifications. Both require the integrity of P-element ends. We have also observed a high frequency of double P elements localized at the original P[w(d1)] insertion site. These double P elements are arranged in nonrandom configurations. We discuss the frequencies and the possible mechanisms leading to the various types of derivatives, in light of the current models for P excision and transposition. We propose that the P transposase induces mainly localized events. Some of these could result from frequent changes of template during gap-repair DNA synthesis, and/or from abortive transposition.
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- 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]
- Chédin F., Dervyn E., Dervyn R., Ehrlich S. D., Noirot P. Frequency of deletion formation decreases exponentially with distance between short direct repeats. Mol Microbiol. 1994 May;12(4):561–569. doi: 10.1111/j.1365-2958.1994.tb01042.x. [DOI] [PubMed] [Google Scholar]
- Coen D. P element regulatory products enhance zeste repression of a P[white duplicated] transgene in Drosophila melanogaster. Genetics. 1990 Dec;126(4):949–960. doi: 10.1093/genetics/126.4.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coen E. S., Carpenter R. A semi-dominant allele, niv-525, acts in trans to inhibit expression of its wild-type homologue in Antirrhinum majus. EMBO J. 1988 Apr;7(4):877–883. doi: 10.1002/j.1460-2075.1988.tb02891.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Dorer D. R., Henikoff S. Expansions of transgene repeats cause heterochromatin formation and gene silencing in Drosophila. Cell. 1994 Jul 1;77(7):993–1002. doi: 10.1016/0092-8674(94)90439-1. [DOI] [PubMed] [Google Scholar]
- Engels W. R. A trans-acting product needed for P factor transposition in Drosophila. Science. 1984 Dec 7;226(4679):1194–1196. doi: 10.1126/science.6095450. [DOI] [PubMed] [Google Scholar]
- 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]
- Gonzy-Tréboul G., Lepesant J. A., Deutsch J. Enhancer-trap targeting at the Broad-Complex locus of Drosophila melanogaster. Genes Dev. 1995 May 1;9(9):1137–1148. doi: 10.1101/gad.9.9.1137. [DOI] [PubMed] [Google Scholar]
- Hama C., Ali Z., Kornberg T. B. Region-specific recombination and expression are directed by portions of the Drosophila engrailed promoter. Genes Dev. 1990 Jul;4(7):1079–1093. doi: 10.1101/gad.4.7.1079. [DOI] [PubMed] [Google Scholar]
- Hawley R. S., Steuber R. A., Marcus C. H., Sohn R., Baronas D. M., Cameron M. L., Zitron A. E., Chase J. W. Molecular analysis of an unstable P element insertion at the singed locus of Drosophila melanogaster: evidence for intracistronic transposition of a P element. Genetics. 1988 May;119(1):85–94. doi: 10.1093/genetics/119.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Kassis J. A., Noll E., VanSickle E. P., Odenwald W. F., Perrimon N. Altering the insertional specificity of a Drosophila transposable element. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1919–1923. doi: 10.1073/pnas.89.5.1919. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Lemaitre B., Coen D. P regulatory products repress in vivo the P promoter activity in P-lacZ fusion genes. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4419–4423. doi: 10.1073/pnas.88.10.4419. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levis R., Hazelrigg T., Rubin G. M. Effects of genomic position on the expression of transduced copies of the white gene of Drosophila. Science. 1985 Aug 9;229(4713):558–561. doi: 10.1126/science.2992080. [DOI] [PubMed] [Google Scholar]
- Levis R., Hazelrigg T., Rubin G. M. Separable cis-acting control elements for expression of the white gene of Drosophila. EMBO J. 1985 Dec 16;4(13A):3489–3499. doi: 10.1002/j.1460-2075.1985.tb04108.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lin F. L., Sperle K., Sternberg N. Model for homologous recombination during transfer of DNA into mouse L cells: role for DNA ends in the recombination process. Mol Cell Biol. 1984 Jun;4(6):1020–1034. doi: 10.1128/mcb.4.6.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Misra S., Rio D. C. Cytotype control of Drosophila P element transposition: the 66 kd protein is a repressor of transposase activity. Cell. 1990 Jul 27;62(2):269–284. doi: 10.1016/0092-8674(90)90365-l. [DOI] [PubMed] [Google Scholar]
- Mullins M. C., Rio D. C., Rubin G. M. cis-acting DNA sequence requirements for P-element transposition. Genes Dev. 1989 May;3(5):729–738. doi: 10.1101/gad.3.5.729. [DOI] [PubMed] [Google Scholar]
- Nassif N., Penney J., Pal S., Engels W. R., Gloor G. B. Efficient copying of nonhomologous sequences from ectopic sites via P-element-induced gap repair. Mol Cell Biol. 1994 Mar;14(3):1613–1625. doi: 10.1128/mcb.14.3.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nitasaka E., Mukai T., Yamazaki T. Repressor of P elements in Drosophila melanogaster: Cytotype determination by a defective P element carrying only open reading frames 0 through 2. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7605–7608. doi: 10.1073/pnas.84.21.7605. [DOI] [PMC free article] [PubMed] [Google Scholar]
- O'Hare K., Driver A., McGrath S., Johnson-Schiltz D. M. Distribution and structure of cloned P elements from the Drosophila melanogaster P strain pi 2. Genet Res. 1992 Aug;60(1):33–41. doi: 10.1017/s0016672300030640. [DOI] [PubMed] [Google Scholar]
- O'Hare K., Murphy C., Levis R., Rubin G. M. DNA sequence of the white locus of Drosophila melanogaster. J Mol Biol. 1984 Dec 15;180(3):437–455. doi: 10.1016/0022-2836(84)90021-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Peterson K. M., Davis P. S., Judd B. H. The determined state of white expression in the Drosophila eye is modified by zeste1 in the wzm family of mutants. Mol Gen Genet. 1994 Mar;242(6):717–726. doi: 10.1007/BF00283427. [DOI] [PubMed] [Google Scholar]
- Pirrotta V., Bröckl C. Transcription of the Drosophila white locus and some of its mutants. EMBO J. 1984 Mar;3(3):563–568. doi: 10.1002/j.1460-2075.1984.tb01847.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pirrotta V., Steller H., Bozzetti M. P. Multiple upstream regulatory elements control the expression of the Drosophila white gene. EMBO J. 1985 Dec 16;4(13A):3501–3508. doi: 10.1002/j.1460-2075.1985.tb04109.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pirrotta V. The genetics and molecular biology of zeste in Drosophila melanogaster. Adv Genet. 1991;29:301–348. doi: 10.1016/s0065-2660(08)60110-8. [DOI] [PubMed] [Google Scholar]
- 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]
- Pâques F., Wegnez M. Deletions and amplifications of tandemly arranged ribosomal 5S genes internal to a P element occur at a high rate in a dysgenic context. Genetics. 1993 Oct;135(2):469–476. doi: 10.1093/genetics/135.2.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Qian S., Varjavand B., Pirrotta V. Molecular analysis of the zeste-white interaction reveals a promoter-proximal element essential for distant enhancer-promoter communication. Genetics. 1992 May;131(1):79–90. doi: 10.1093/genetics/131.1.79. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rio D. C., Laski F. A., Rubin G. M. Identification and immunochemical analysis of biologically active Drosophila P element transposase. Cell. 1986 Jan 17;44(1):21–32. doi: 10.1016/0092-8674(86)90481-2. [DOI] [PubMed] [Google Scholar]
- 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]
- Roiha H., Rubin G. M., O'Hare K. P element insertions and rearrangements at the singed locus of Drosophila melanogaster. Genetics. 1988 May;119(1):75–83. doi: 10.1093/genetics/119.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rubin G. M., Hazelrigg T., Karess R. E., Laski F. A., Laverty T., Levis R., Rio D. C., Spencer F. A., Zuker C. S. Germ line specificity of P-element transposition and some novel patterns of expression of transduced copies of the white gene. Cold Spring Harb Symp Quant Biol. 1985;50:329–335. doi: 10.1101/sqb.1985.050.01.042. [DOI] [PubMed] [Google Scholar]
- Rubin G. M., Kidwell M. G., Bingham P. M. The molecular basis of P-M hybrid dysgenesis: the nature of induced mutations. Cell. 1982 Jul;29(3):987–994. doi: 10.1016/0092-8674(82)90462-7. [DOI] [PubMed] [Google Scholar]
- Sacramento D., Bourhy H., Tordo N. PCR technique as an alternative method for diagnosis and molecular epidemiology of rabies virus. Mol Cell Probes. 1991 Jun;5(3):229–240. doi: 10.1016/0890-8508(91)90045-l. [DOI] [PubMed] [Google Scholar]
- Spradling A. C., Rubin G. M. Transposition of cloned P elements into Drosophila germ line chromosomes. Science. 1982 Oct 22;218(4570):341–347. doi: 10.1126/science.6289435. [DOI] [PubMed] [Google Scholar]
- Sugawara N., Haber J. E. Characterization of double-strand break-induced recombination: homology requirements and single-stranded DNA formation. Mol Cell Biol. 1992 Feb;12(2):563–575. doi: 10.1128/mcb.12.2.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Thompson-Stewart D., Karpen G. H., Spradling A. C. A transposable element can drive the concerted evolution of tandemly repetitious DNA. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9042–9046. doi: 10.1073/pnas.91.19.9042. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tower J., Karpen G. H., Craig N., Spradling A. C. Preferential transposition of Drosophila P elements to nearby chromosomal sites. Genetics. 1993 Feb;133(2):347–359. doi: 10.1093/genetics/133.2.347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhang P., Spradling A. C. Efficient and dispersed local P element transposition from Drosophila females. Genetics. 1993 Feb;133(2):361–373. doi: 10.1093/genetics/133.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]