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. 1994 Oct;14(10):6689–6695. doi: 10.1128/mcb.14.10.6689

Integration of a vector containing a repetitive LINE-1 element in the human genome.

M Richard 1, A Belmaaza 1, N Gusew 1, J C Wallenburg 1, P Chartrand 1
PMCID: PMC359199  PMID: 7935388

Abstract

Mammalian cells contain numerous nonallelic repeated sequences, such as multicopy genes, gene families, and repeated elements. One common feature of nonallelic repeated sequences is that they are homeologous (not perfectly identical). Our laboratory has been studying recombination between homeologous sequences by using LINE-1 (L1) elements as substrates. We showed previously that an exogenous L1 element could readily acquire endogenous L1 sequences by nonreciprocal homologous recombination. In the study presented here, we have investigated the propensity of exogenous L1 elements to be involved in a reciprocal process, namely, crossing-overs. This would result in the integration of the exogenous L1 element into an endogenous L1 element. Of over 400 distinct integration events analyzed, only 2% involved homologous recombination between exogenous and endogenous L1 elements. These homologous recombination events were imprecise, with the integrated vector being flanked by one homologous and one illegitimate junction. This type of structure is not consistent with classical crossing-overs that would result in two homologous junctions but rather is consistent with one-sided homologous recombination followed by illegitimate integration. Contrary to what has been found for reciprocal homologous integration, the degree of homology between the exogenous and endogenous L1 elements did not seem to play an important role in the choice of recombination partners. These results suggest that although exogenous and endogenous L1 elements are capable of homologous recombination, this seldom leads to crossing-overs. This observation could have implications for the stability of mammalian genomes.

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Selected References

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

  1. Adair G. M., Nairn R. S., Wilson J. H., Seidman M. M., Brotherman K. A., MacKinnon C., Scheerer J. B. Targeted homologous recombination at the endogenous adenine phosphoribosyltransferase locus in Chinese hamster cells. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4574–4578. doi: 10.1073/pnas.86.12.4574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aratani Y., Okazaki R., Koyama H. End extension repair of introduced targeting vectors mediated by homologous recombination in mammalian cells. Nucleic Acids Res. 1992 Sep 25;20(18):4795–4801. doi: 10.1093/nar/20.18.4795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Belmaaza A., Chartrand P. One-sided invasion events in homologous recombination at double-strand breaks. Mutat Res. 1994 May;314(3):199–208. doi: 10.1016/0921-8777(94)90065-5. [DOI] [PubMed] [Google Scholar]
  4. Belmaaza A., Wallenburg J. C., Brouillette S., Gusew N., Chartrand P. Genetic exchange between endogenous and exogenous LINE-1 repetitive elements in mouse cells. Nucleic Acids Res. 1990 Nov 11;18(21):6385–6391. doi: 10.1093/nar/18.21.6385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berinstein N., Pennell N., Ottaway C. A., Shulman M. J. Gene replacement with one-sided homologous recombination. Mol Cell Biol. 1992 Jan;12(1):360–367. doi: 10.1128/mcb.12.1.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chu G., Hayakawa H., Berg P. Electroporation for the efficient transfection of mammalian cells with DNA. Nucleic Acids Res. 1987 Feb 11;15(3):1311–1326. doi: 10.1093/nar/15.3.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Colbere-Garapin F., Horodniceanu F., Kourilsky P., Garapin A. C. Cloning of the herpes simplex virus type 1 thymidine kinase gene in E. coli K 12: a selective marker for gene transfer into animal cells. Dev Biol Stand. 1980;46:75–82. [PubMed] [Google Scholar]
  8. Deng C., Capecchi M. R. Reexamination of gene targeting frequency as a function of the extent of homology between the targeting vector and the target locus. Mol Cell Biol. 1992 Aug;12(8):3365–3371. doi: 10.1128/mcb.12.8.3365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dover G. Molecular drive: a cohesive mode of species evolution. Nature. 1982 Sep 9;299(5879):111–117. doi: 10.1038/299111a0. [DOI] [PubMed] [Google Scholar]
  10. Ellis J., Bernstein A. Gene targeting with retroviral vectors: recombination by gene conversion into regions of nonhomology. Mol Cell Biol. 1989 Apr;9(4):1621–1627. doi: 10.1128/mcb.9.4.1621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grimaldi G., Skowronski J., Singer M. F. Defining the beginning and end of KpnI family segments. EMBO J. 1984 Aug;3(8):1753–1759. doi: 10.1002/j.1460-2075.1984.tb02042.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Higashi Y., Tanae A., Inoue H., Fujii-Kuriyama Y. Evidence for frequent gene conversion in the steroid 21-hydroxylase P-450(C21) gene: implications for steroid 21-hydroxylase deficiency. Am J Hum Genet. 1988 Jan;42(1):17–25. [PMC free article] [PubMed] [Google Scholar]
  13. Hu X. Y., Ray P. N., Worton R. G. Mechanisms of tandem duplication in the Duchenne muscular dystrophy gene include both homologous and nonhomologous intrachromosomal recombination. EMBO J. 1991 Sep;10(9):2471–2477. doi: 10.1002/j.1460-2075.1991.tb07786.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hwu H. R., Roberts J. W., Davidson E. H., Britten R. J. Insertion and/or deletion of many repeated DNA sequences in human and higher ape evolution. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3875–3879. doi: 10.1073/pnas.83.11.3875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jurka J. Subfamily structure and evolution of the human L1 family of repetitive sequences. J Mol Evol. 1989 Dec;29(6):496–503. doi: 10.1007/BF02602921. [DOI] [PubMed] [Google Scholar]
  16. Knight K. L. Restricted VH gene usage and generation of antibody diversity in rabbit. Annu Rev Immunol. 1992;10:593–616. doi: 10.1146/annurev.iy.10.040192.003113. [DOI] [PubMed] [Google Scholar]
  17. Lehrman M. A., Goldstein J. L., Russell D. W., Brown M. S. Duplication of seven exons in LDL receptor gene caused by Alu-Alu recombination in a subject with familial hypercholesterolemia. Cell. 1987 Mar 13;48(5):827–835. doi: 10.1016/0092-8674(87)90079-1. [DOI] [PubMed] [Google Scholar]
  18. Metzenberg A. B., Wurzer G., Huisman T. H., Smithies O. Homology requirements for unequal crossing over in humans. Genetics. 1991 May;128(1):143–161. doi: 10.1093/genetics/128.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Miller C. C., McPheat J. C., Potts W. J. Targeted integration of the Ren-1D locus in mouse embryonic stem cells. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5020–5024. doi: 10.1073/pnas.89.11.5020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Roeder G. S. Chromosome synapsis and genetic recombination: their roles in meiotic chromosome segregation. Trends Genet. 1990 Dec;6(12):385–389. doi: 10.1016/0168-9525(90)90297-j. [DOI] [PubMed] [Google Scholar]
  21. Rouyer F., Simmler M. C., Page D. C., Weissenbach J. A sex chromosome rearrangement in a human XX male caused by Alu-Alu recombination. Cell. 1987 Nov 6;51(3):417–425. doi: 10.1016/0092-8674(87)90637-4. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Scott A. F., Schmeckpeper B. J., Abdelrazik M., Comey C. T., O'Hara B., Rossiter J. P., Cooley T., Heath P., Smith K. D., Margolet L. Origin of the human L1 elements: proposed progenitor genes deduced from a consensus DNA sequence. Genomics. 1987 Oct;1(2):113–125. doi: 10.1016/0888-7543(87)90003-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Skowronski J., Fanning T. G., Singer M. F. Unit-length line-1 transcripts in human teratocarcinoma cells. Mol Cell Biol. 1988 Apr;8(4):1385–1397. doi: 10.1128/mcb.8.4.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  26. 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]
  27. Thomas K. R., Capecchi M. R. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell. 1987 Nov 6;51(3):503–512. doi: 10.1016/0092-8674(87)90646-5. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Wheeler C. J., Maloney D., Fogel S., Goodenow R. S. Microconversion between murine H-2 genes integrated into yeast. Nature. 1990 Sep 13;347(6289):192–194. doi: 10.1038/347192a0. [DOI] [PubMed] [Google Scholar]
  30. Yenofsky R. L., Fine M., Pellow J. W. A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure. Proc Natl Acad Sci U S A. 1990 May;87(9):3435–3439. doi: 10.1073/pnas.87.9.3435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zheng H., Wilson J. H. Gene targeting in normal and amplified cell lines. Nature. 1990 Mar 8;344(6262):170–173. doi: 10.1038/344170a0. [DOI] [PubMed] [Google Scholar]

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