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. 1992 Aug 25;20(16):4269–4274. doi: 10.1093/nar/20.16.4269

Molecular analysis of DNA junctions produced by illegitimate recombination in human cells.

A Stary 1, A Sarasin 1
PMCID: PMC334135  PMID: 1324477

Abstract

In a human HeLa derived-cell line carrying permanently a single integrated copy of an SV40 shuttle vector, the transient expression of the SV40 T-antigen led to the production of heterogeneous populations of circular DNA molecules which retained both integrated vector and its surrounding cellular sequences. Comparison between the integrated copy and the linear maps of 80 different plasmids rescued in bacteria suggested that the formation of circular DNA was the result of bidirectional replication from the SV40 origin of replication followed by a single intramolecular joining leading to the cyclization of the replicated molecules. Sequence analysis of 45 recombinational junctions demonstrated that the cyclization occurred via illegitimate recombination process which did not require preferential nucleotide sequence at the joining sites. However, extensive characterization of recombination junctions revealed that the sequences involved in the recombination at each side of the SV40 origin of replication were not randomly distributed, suggesting the presence of regions which were more prone to be involved in the illegitimate recombination process in human cells. Search of common features usually implied in illegitimate recombination in mammalian cells revealed some association of these regions with palindromes, A + T-rich DNA segments, alternating purine/pyrimidine sequences and Alu family repeats.

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Selected References

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

  1. Bishop J. M. Molecular themes in oncogenesis. Cell. 1991 Jan 25;64(2):235–248. doi: 10.1016/0092-8674(91)90636-d. [DOI] [PubMed] [Google Scholar]
  2. Borowiec J. A., Dean F. B., Bullock P. A., Hurwitz J. Binding and unwinding--how T antigen engages the SV40 origin of DNA replication. Cell. 1990 Jan 26;60(2):181–184. doi: 10.1016/0092-8674(90)90730-3. [DOI] [PubMed] [Google Scholar]
  3. Botchan M., Stringer J., Mitchison T., Sambrook J. Integration and excision of SV40 DNA from the chromosome of a transformed cell. Cell. 1980 May;20(1):143–152. doi: 10.1016/0092-8674(80)90242-1. [DOI] [PubMed] [Google Scholar]
  4. Bullock P., Forrester W., Botchan M. DNA sequence studies of simian virus 40 chromosomal excision and integration in rat cells. J Mol Biol. 1984 Mar 25;174(1):55–84. doi: 10.1016/0022-2836(84)90365-6. [DOI] [PubMed] [Google Scholar]
  5. Gerard R. D., Gluzman Y. New host cell system for regulated simian virus 40 DNA replication. Mol Cell Biol. 1985 Nov;5(11):3231–3240. doi: 10.1128/mcb.5.11.3231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gutai M. W., Nathans D. Evolutionary variants of simian virus 40: Cellular DNA sequences and sequences at recombinant joints of substituted variants. J Mol Biol. 1978 Dec 5;126(2):275–288. doi: 10.1016/0022-2836(78)90363-7. [DOI] [PubMed] [Google Scholar]
  7. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  8. Heinzel S. S., Krysan P. J., Calos M. P., DuBridge R. B. Use of simian virus 40 replication to amplify Epstein-Barr virus shuttle vectors in human cells. J Virol. 1988 Oct;62(10):3738–3746. doi: 10.1128/jvi.62.10.3738-3746.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hyrien O., Debatisse M., Buttin G., de Saint Vincent B. R. A hotspot for novel amplification joints in a mosaic of Alu-like repeats and palindromic A + T-rich DNA. EMBO J. 1987 Aug;6(8):2401–2408. doi: 10.1002/j.1460-2075.1987.tb02518.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Konopka A. K. Compilation of DNA strand exchange sites for non-homologous recombination in somatic cells. Nucleic Acids Res. 1988 Mar 11;16(5):1739–1758. doi: 10.1093/nar/16.5.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kowalski D., Natale D. A., Eddy M. J. Stable DNA unwinding, not "breathing," accounts for single-strand-specific nuclease hypersensitivity of specific A+T-rich sequences. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9464–9468. doi: 10.1073/pnas.85.24.9464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Menck C. F., Sarasin A., James M. R. SV40-based Escherichia coli shuttle vectors infectious for monkey cells. Gene. 1987;53(1):21–29. doi: 10.1016/0378-1119(87)90089-8. [DOI] [PubMed] [Google Scholar]
  13. Miller J., Bullock P., Botchan M. Simian virus 40 T antigen is required for viral excision from chromosomes. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7534–7538. doi: 10.1073/pnas.81.23.7534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nalbantoglu J., Hartley D., Phear G., Tear G., Meuth M. Spontaneous deletion formation at the aprt locus of hamster cells: the presence of short sequence homologies and dyad symmetries at deletion termini. EMBO J. 1986 Jun;5(6):1199–1204. doi: 10.1002/j.1460-2075.1986.tb04347.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Roth D. B., Chang X. B., Wilson J. H. Comparison of filler DNA at immune, nonimmune, and oncogenic rearrangements suggests multiple mechanisms of formation. Mol Cell Biol. 1989 Jul;9(7):3049–3057. doi: 10.1128/mcb.9.7.3049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schiedner G., Wessel R., Scheffner M., Stahl H. Renaturation and DNA looping promoted by the SV40 large tumour antigen. EMBO J. 1990 Sep;9(9):2937–2943. doi: 10.1002/j.1460-2075.1990.tb07485.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schimke R. T. Gene amplification in cultured cells. J Biol Chem. 1988 May 5;263(13):5989–5992. [PubMed] [Google Scholar]
  18. Stark G. R., Debatisse M., Giulotto E., Wahl G. M. Recent progress in understanding mechanisms of mammalian DNA amplification. Cell. 1989 Jun 16;57(6):901–908. doi: 10.1016/0092-8674(89)90328-0. [DOI] [PubMed] [Google Scholar]
  19. Stary A., James M. R., Sarasin A. High recombination rate of an Epstein-Barr virus-simian virus 40 hybrid shuttle vector in human cells. J Virol. 1989 Sep;63(9):3837–3843. doi: 10.1128/jvi.63.9.3837-3843.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wahl G. M. The importance of circular DNA in mammalian gene amplification. Cancer Res. 1989 Mar 15;49(6):1333–1340. [PubMed] [Google Scholar]
  21. Wessel R., Schweizer J., Stahl H. Simian virus 40 T-antigen DNA helicase is a hexamer which forms a binary complex during bidirectional unwinding from the viral origin of DNA replication. J Virol. 1992 Feb;66(2):804–815. doi: 10.1128/jvi.66.2.804-815.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wigler M., Pellicer A., Silverstein S., Axel R. Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell. 1978 Jul;14(3):725–731. doi: 10.1016/0092-8674(78)90254-4. [DOI] [PubMed] [Google Scholar]
  23. Zerbib D., Gamas P., Chandler M., Prentki P., Bass S., Galas D. Specificity of insertion of IS1. J Mol Biol. 1985 Oct 5;185(3):517–524. doi: 10.1016/0022-2836(85)90068-3. [DOI] [PubMed] [Google Scholar]

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