Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1991 Nov 11;19(21):5943–5947. doi: 10.1093/nar/19.21.5943

Stimulation of intrachromosomal homologous recombination in mammalian cells by an inhibitor of poly(ADP-ribosylation).

A S Waldman 1, B C Waldman 1
PMCID: PMC329051  PMID: 1945881

Abstract

We determined the effect of 3-methoxybenzamide (3-MB), a competitive inhibitor of poly(ADP-ribose) polymerase (E.C. 2.4.2.30), on intrachromosomal homologous recombination in mouse Ltk- cells. We used a cell line that contained in its genome two defective Herpes thymidine kinase (tk) genes as closely linked direct repeats. Intrachromosomal homologous recombination events were monitored by selecting for tk-positive segregants that arose during propagation of the cells and recombination rates were determined by fluctuation analysis. We found that growth of cells in the continuous presence of 2mM 3-MB increased intrachromosomal recombination between 3 and 4-fold. Growth of cells in the presence of 2mM m-anisic acid, a non-inhibitory analog of 3-MB, had no effect on intrachromosomal recombination rates. Additionally, we found that 3-MB increased both gene conversions and crossovers to similar extents, adding to the evidence that these two types of intrachromosomal rearrangements share a common pathway. These findings contrast with our previous studies [Waldman, B.C. and Waldman, A.S. (1990) Nucleic Acids Res., 18, 5981-5988] in which we determined that 3-MB inhibits illegitimate recombination and has no effect on extrachromosomal homologous recombination in mouse Ltk- cells. An hypothesis is offered that explains the influence of 3-MB on different recombination pathways in mammalian cells in terms of the role that poly(ADP-ribosylation) plays in DNA break-repair.

Full text

PDF
5943

Images in this article

5946Image
on p.5946

Selected References

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

  1. Arlett C. F., Lehmann A. R. Human disorders showing increased sensitivity to the induction of genetic damage. Annu Rev Genet. 1978;12:95–115. doi: 10.1146/annurev.ge.12.120178.000523. [DOI] [PubMed] [Google Scholar]
  2. Benjamin R. C., Gill D. M. Poly(ADP-ribose) synthesis in vitro programmed by damaged DNA. A comparison of DNA molecules containing different types of strand breaks. J Biol Chem. 1980 Nov 10;255(21):10502–10508. [PubMed] [Google Scholar]
  3. Bollag R. J., Liskay R. M. Conservative intrachromosomal recombination between inverted repeats in mouse cells: association between reciprocal exchange and gene conversion. Genetics. 1988 May;119(1):161–169. doi: 10.1093/genetics/119.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bollag R. J., Waldman A. S., Liskay R. M. Homologous recombination in mammalian cells. Annu Rev Genet. 1989;23:199–225. doi: 10.1146/annurev.ge.23.120189.001215. [DOI] [PubMed] [Google Scholar]
  5. Capecchi M. R. Altering the genome by homologous recombination. Science. 1989 Jun 16;244(4910):1288–1292. doi: 10.1126/science.2660260. [DOI] [PubMed] [Google Scholar]
  6. Capecchi M. R. The new mouse genetics: altering the genome by gene targeting. Trends Genet. 1989 Mar;5(3):70–76. doi: 10.1016/0168-9525(89)90029-2. [DOI] [PubMed] [Google Scholar]
  7. Capizzi R. L., Jameson J. W. A table for the estimation of the spontaneous mutation rate of cells in culture. Mutat Res. 1973 Jan;17(1):147–148. doi: 10.1016/0027-5107(73)90265-0. [DOI] [PubMed] [Google Scholar]
  8. Creissen D., Shall S. Regulation of DNA ligase activity by poly(ADP-ribose). Nature. 1982 Mar 18;296(5854):271–272. doi: 10.1038/296271a0. [DOI] [PubMed] [Google Scholar]
  9. Farzaneh F., Panayotou G. N., Bowler L. D., Hardas B. D., Broom T., Walther C., Shall S. ADP-ribosylation is involved in the integration of foreign DNA into the mammalian cell genome. Nucleic Acids Res. 1988 Dec 9;16(23):11319–11326. doi: 10.1093/nar/16.23.11319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Frohman M. A., Martin G. R. Cut, paste, and save: new approaches to altering specific genes in mice. Cell. 1989 Jan 27;56(2):145–147. doi: 10.1016/0092-8674(89)90887-8. [DOI] [PubMed] [Google Scholar]
  11. Liskay R. M., Stachelek J. L., Letsou A. Homologous recombination between repeated chromosomal sequences in mouse cells. Cold Spring Harb Symp Quant Biol. 1984;49:183–189. doi: 10.1101/sqb.1984.049.01.021. [DOI] [PubMed] [Google Scholar]
  12. Luria S. E., Delbrück M. Mutations of Bacteria from Virus Sensitivity to Virus Resistance. Genetics. 1943 Nov;28(6):491–511. doi: 10.1093/genetics/28.6.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nakayasu M., Shima H., Aonuma S., Nakagama H., Nagao M., Sugimura T. Deletion of transfected oncogenes from NIH 3T3 transformants by inhibitors of poly(ADP-ribose) polymerase. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9066–9070. doi: 10.1073/pnas.85.23.9066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Orr-Weaver T. L., Szostak J. W. Fungal recombination. Microbiol Rev. 1985 Mar;49(1):33–58. doi: 10.1128/mr.49.1.33-58.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Purnell M. R., Whish W. J. Novel inhibitors of poly(ADP-ribose) synthetase. Biochem J. 1980 Mar 1;185(3):775–777. doi: 10.1042/bj1850775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wagner M. J., Sharp J. A., Summers W. C. Nucleotide sequence of the thymidine kinase gene of herpes simplex virus type 1. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1441–1445. doi: 10.1073/pnas.78.3.1441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Waldman A. S., Liskay R. M. Differential effects of base-pair mismatch on intrachromosomal versus extrachromosomal recombination in mouse cells. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5340–5344. doi: 10.1073/pnas.84.15.5340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Waldman B. C., Waldman A. S. Illegitimate and homologous recombination in mammalian cells: differential sensitivity to an inhibitor of poly(ADP-ribosylation). Nucleic Acids Res. 1990 Oct 25;18(20):5981–5988. doi: 10.1093/nar/18.20.5981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wang Y. Y., Maher V. M., Liskay R. M., McCormick J. J. Carcinogens can induce homologous recombination between duplicated chromosomal sequences in mouse L cells. Mol Cell Biol. 1988 Jan;8(1):196–202. doi: 10.1128/mcb.8.1.196. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES