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. 1990 May;64(5):2327–2336. doi: 10.1128/jvi.64.5.2327-2336.1990

Preferred crossover sites on polyomavirus DNA.

P Bourgaux 1, D Gendron 1, D Bourgaux-Ramoisy 1
PMCID: PMC249394  PMID: 2157889

Abstract

RmI is a hybrid replicon consisting of polyomavirus (Py) and mouse sequences that yields unit-length polyomavirus DNA via recombination between two directly repeated viral sequences of 182 base pairs (S repeats). To define the contribution of the S repeats in this intramolecular recombination, we derived from RmI a series of replicons containing the original S repeats as well as additional direct viral repeats which were 1 to 2 kilobases in length (L repeats). After mouse 3T6 cells were transfected with these constructs, recombination products that displayed the physical properties of homologous recombinants were detected. The structures of these recombinants indicated that whereas repeat length influences the likelihood of recombination, crossover occurs preferentially near the S repeats, provided that one of them is proximal to the viral origin of replication. This finding suggests that recombination near the S repeats depends on a process initiated near the viral origin of replication.

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Selected References

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

  1. Basilico C., Gattoni S., Zouzias D., Valle G. D. Loss of integrated viral DNA sequences in polyomatransformed cells is associated with an active viral A function. Cell. 1979 Jul;17(3):645–659. doi: 10.1016/0092-8674(79)90272-1. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Botchan M., Topp W., Sambrook J. Studies on simian virus 40 excision from cellular chromosomes. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):709–719. doi: 10.1101/sqb.1979.043.01.079. [DOI] [PubMed] [Google Scholar]
  4. Bourgaux-Ramoisy D., Gendron D., Chartrand P., Bourgaux P. An excision event that may depend on patchy homology for site specificity. Mol Cell Biol. 1986 Jul;6(7):2727–2730. doi: 10.1128/mcb.6.7.2727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bourgaux P., Delbecchi L., Yu K. K., Herring E., Bourgaux-Ramoisy D. A mouse embryo cell line carrying an inducible, temperature-sensitive, polyoma virus genome. Virology. 1978 Jul 15;88(2):348–360. doi: 10.1016/0042-6822(78)90291-x. [DOI] [PubMed] [Google Scholar]
  6. Bourgaux P., Sylla B. S., Chartrand P. Excision of polyoma virus DNA from that of a transformed mouse cell: identification of a hybrid molecule with direct and inverted repeat sequences at the viral-cellular joints. Virology. 1982 Oct 15;122(1):84–97. doi: 10.1016/0042-6822(82)90379-8. [DOI] [PubMed] [Google Scholar]
  7. Brouillette S., Chartrand P. Intermolecular recombination assay for mammalian cells that produces recombinants carrying both homologous and nonhomologous junctions. Mol Cell Biol. 1987 Jun;7(6):2248–2255. doi: 10.1128/mcb.7.6.2248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chakrabarti S., Joffe S., Seidman M. M. Recombination and deletion of sequences in shuttle vector plasmids in mammalian cells. Mol Cell Biol. 1985 Sep;5(9):2265–2271. doi: 10.1128/mcb.5.9.2265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Colantuoni V., Dailey L., Valle G. D., Basilico C. Requirements for excision and amplification of integrated viral DNA molecules in polyoma virus-transformed cells. J Virol. 1982 Aug;43(2):617–628. doi: 10.1128/jvi.43.2.617-628.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Delbecchi L., Gendron D., Bourgaux P. Inducible permissive cells transformed by a temperature-sensitive polyoma virus: superinfection does not allow excision of the resident viral genome. J Virol. 1981 Jul;39(1):196–206. doi: 10.1128/jvi.39.1.196-206.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. FRIED M. CELL-TRANSFORMING ABILITY OF A TEMPERATURE-SENSITIVE MUTANT OF POLYOMA VIRUS. Proc Natl Acad Sci U S A. 1965 Mar;53:486–491. doi: 10.1073/pnas.53.3.486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gattoni S., Colantuoni V., Basilico C. Relationship between integrated and nonintegrated viral DNA in rat cells transformed by polyoma virus. J Virol. 1980 Jun;34(3):615–626. doi: 10.1128/jvi.34.3.615-626.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gendron D., Delbecchi L., Bourgaux-Ramoisy D., Bourgaux P. A substitution in a nonconserved region of polyomavirus large T antigen which causes a thermosensitive mutation. Virology. 1988 Jul;165(1):165–171. doi: 10.1016/0042-6822(88)90669-1. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Hinnen A., Hicks J. B., Fink G. R. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. doi: 10.1073/pnas.75.4.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  17. Holliday R. Molecular aspects of genetic exchange and gene conversion. Genetics. 1974 Sep;78(1):273–287. doi: 10.1093/genetics/78.1.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jasin M., de Villiers J., Weber F., Schaffner W. High frequency of homologous recombination in mammalian cells between endogenous and introduced SV40 genomes. Cell. 1985 Dec;43(3 Pt 2):695–703. doi: 10.1016/0092-8674(85)90242-9. [DOI] [PubMed] [Google Scholar]
  19. Lania L., Boast S., Fried M. Excision of polyoma virus genomes from chromosomal DNA by homologous recombination. Nature. 1982 Jan 28;295(5847):349–350. doi: 10.1038/295349a0. [DOI] [PubMed] [Google Scholar]
  20. Lin F. L., Sperle K., Sternberg N. Recombination in mouse L cells between DNA introduced into cells and homologous chromosomal sequences. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1391–1395. doi: 10.1073/pnas.82.5.1391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Meselson M. S., Radding C. M. A general model for genetic recombination. Proc Natl Acad Sci U S A. 1975 Jan;72(1):358–361. doi: 10.1073/pnas.72.1.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. doi: 10.1073/pnas.78.10.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Orr-Weaver T. L., Szostak J. W. Yeast recombination: the association between double-strand gap repair and crossing-over. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4417–4421. doi: 10.1073/pnas.80.14.4417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pellegrini S., Dailey L., Basilico C. Amplification and excision of integrated polyoma DNA sequences require a functional origin of replication. Cell. 1984 Apr;36(4):943–949. doi: 10.1016/0092-8674(84)90044-8. [DOI] [PubMed] [Google Scholar]
  26. Piché A., Bourgaux P. Resolution of a polyomavirus-mouse hybrid replicon: release of genomic viral DNA. J Virol. 1987 Mar;61(3):840–844. doi: 10.1128/jvi.61.3.840-844.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Piché A., Bourgaux P. Resolution of a polyomavirus-mouse hybrid replicon: viral function required for recombination. J Virol. 1987 Mar;61(3):845–850. doi: 10.1128/jvi.61.3.845-850.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rao B. S., Martin R. G. DpnI assay for DNA replication in animal cells: enzyme-resistant material can result from factors not related to replication. Nucleic Acids Res. 1988 May 11;16(9):4171–4171. doi: 10.1093/nar/16.9.4171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Roth D. B., Wilson J. H. Relative rates of homologous and nonhomologous recombination in transfected DNA. Proc Natl Acad Sci U S A. 1985 May;82(10):3355–3359. doi: 10.1073/pnas.82.10.3355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rubnitz J., Subramani S. Rapid assay for extrachromosomal homologous recombination in monkey cells. Mol Cell Biol. 1985 Mar;5(3):529–537. doi: 10.1128/mcb.5.3.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rubnitz J., Subramani S. The minimum amount of homology required for homologous recombination in mammalian cells. Mol Cell Biol. 1984 Nov;4(11):2253–2258. doi: 10.1128/mcb.4.11.2253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  33. Smith A. J., Berg P. Homologous recombination between defective neo genes in mouse 3T6 cells. Cold Spring Harb Symp Quant Biol. 1984;49:171–181. doi: 10.1101/sqb.1984.049.01.020. [DOI] [PubMed] [Google Scholar]
  34. Smithies O., Gregg R. G., Boggs S. S., Koralewski M. A., Kucherlapati R. S. Insertion of DNA sequences into the human chromosomal beta-globin locus by homologous recombination. Nature. 1985 Sep 19;317(6034):230–234. doi: 10.1038/317230a0. [DOI] [PubMed] [Google Scholar]
  35. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  36. Stringer J. R. Recombination between poly[d(GT).d(CA)] sequences in simian virus 40-infected cultured cells. Mol Cell Biol. 1985 Jun;5(6):1247–1259. doi: 10.1128/mcb.5.6.1247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Subramani S., Berg P. Homologous and nonhomologous recombination in monkey cells. Mol Cell Biol. 1983 Jun;3(6):1040–1052. doi: 10.1128/mcb.3.6.1040. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sussman D. J., Milman G. Short-term, high-efficiency expression of transfected DNA. Mol Cell Biol. 1984 Aug;4(8):1641–1643. doi: 10.1128/mcb.4.8.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sylla B. S., Allard D., Roy G., Bourgaux-Ramoisy D., Bourgaux P. A mouse DNA sequence that mediates integration and excision of polyoma virus DNA. Gene. 1984 Sep;29(3):343–350. doi: 10.1016/0378-1119(84)90063-5. [DOI] [PubMed] [Google Scholar]
  40. Sylla B. S., Huberdeau D., Bourgaux-Ramoisy D., Bourgaux P. Site-specific excision of integrated polyoma DNA. Cell. 1984 Jun;37(2):661–667. doi: 10.1016/0092-8674(84)90398-2. [DOI] [PubMed] [Google Scholar]
  41. Szostak J. W., Orr-Weaver T. L., Rothstein R. J., Stahl F. W. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. doi: 10.1016/0092-8674(83)90331-8. [DOI] [PubMed] [Google Scholar]
  42. Thaler D. S., Stahl F. W. DNA double-chain breaks in recombination of phage lambda and of yeast. Annu Rev Genet. 1988;22:169–197. doi: 10.1146/annurev.ge.22.120188.001125. [DOI] [PubMed] [Google Scholar]
  43. Thomas K. R., Folger K. R., Capecchi M. R. High frequency targeting of genes to specific sites in the mammalian genome. Cell. 1986 Feb 14;44(3):419–428. doi: 10.1016/0092-8674(86)90463-0. [DOI] [PubMed] [Google Scholar]
  44. Wake C. T., Wilson J. H. Defined oligomeric SV40 DNA: a sensitive probe of general recombination in somatic cells. Cell. 1980 Aug;21(1):141–148. doi: 10.1016/0092-8674(80)90121-x. [DOI] [PubMed] [Google Scholar]
  45. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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