Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Sep;71(9):3570–3574. doi: 10.1073/pnas.71.9.3570

Distribution of 5-Bromodeoxyuridine in the DNA of Rat Embryo Cells

Stephen A Schwartz 1,2,3, Werner H Kirsten 1,2,3
PMCID: PMC433816  PMID: 4530322

Abstract

Rat embryo cells were treated with 3H-labeled 5-bromodeoxyuridine (BrdU) in order to determine the nature of substitution and distribution of the analog in DNA. At optimal oncornavirus-inducing doses of BrdU (10-4 M), density gradient centrifugation and DNA·DNA reassociation experiments revealed extensive (>90%) and uniform base substitution in repetitive, intermediate, and unique DNA sequences. Suboptimal doses (10-7 M) of [3H]BrdU resulted in less than 5% thymine replacement and nonrandom labeling of DNA. Treatment of cells with 10-7 M [3H]dT resulted in relatively uniform labeling throughout all DNA sequences. At low concentrations, BrdU was incorporated predominantly within the repetitive and intermediate DNA of rat embryo cells. Moreover, the single-copy DNA sequences were lightly substituted and reassociated extensively only in the presence of excess unlabeled DNA. Such specificity of base substitution may be related to the unusually selective nature of BrdU effects on animal cells.

Keywords: DNA·DNA reassociation

Full text

PDF
3570

Selected References

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

  1. Aaronson S. A., Todaro G. J., Scolnick E. M. Induction of murine C-type viruses from clonal lines of virus-free BALB-3T3 cells. Science. 1971 Oct 8;174(4005):157–159. doi: 10.1126/science.174.4005.157. [DOI] [PubMed] [Google Scholar]
  2. Abbott J., Mayne R., Holtzer H. Inhibition of cartilage development in organ cultures of chick somites by the thymidine analog, 5-bromo-2'-deoxyuridine. Dev Biol. 1972 Jun;28(2):430–442. doi: 10.1016/0012-1606(72)90024-3. [DOI] [PubMed] [Google Scholar]
  3. Baluda M. A., Nayak D. P. DNA complementary to viral RNA in leukemic cells induced by avian myeloblastosis virus. Proc Natl Acad Sci U S A. 1970 Jun;66(2):329–336. doi: 10.1073/pnas.66.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Britten R. J., Davidson E. H. Gene regulation for higher cells: a theory. Science. 1969 Jul 25;165(3891):349–357. doi: 10.1126/science.165.3891.349. [DOI] [PubMed] [Google Scholar]
  5. Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [DOI] [PubMed] [Google Scholar]
  6. Davidson E. H., Hough B. R., Amenson C. S., Britten R. J. General interspersion of repetitive with non-repetitive sequence elements in the DNA of Xenopus. J Mol Biol. 1973 Jun 15;77(1):1–23. doi: 10.1016/0022-2836(73)90359-8. [DOI] [PubMed] [Google Scholar]
  7. Gelb L. D., Martin M. A. Simian virus 40 DNA integration within the genome of virus-transformed mammalian cells. Virology. 1973 Feb;51(2):351–357. doi: 10.1016/0042-6822(73)90434-0. [DOI] [PubMed] [Google Scholar]
  8. Gelb L. D., Milstien J. B., Martin M. A., Aaronson S. A. Characterization of murine leukaemia virus-specific DNA present in normal mouse cells. Nat New Biol. 1973 Jul 18;244(133):76–79. doi: 10.1038/newbio244076a0. [DOI] [PubMed] [Google Scholar]
  9. Gelderman A. H., Rake A. V., Britten R. J. Transcription of nonrepeated DNA in neonatal and fetal mice. Proc Natl Acad Sci U S A. 1971 Jan;68(1):172–176. doi: 10.1073/pnas.68.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldberg R. B., Galau G. A., Britten R. J., Davidson E. H. Nonrepetitive DNA sequence representation in sea urchin embryo messenger RNA. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3516–3520. doi: 10.1073/pnas.70.12.3516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gordon P., Casey J., Rabinowitz M. Characterization of mitochondrial deoxyribonucleic acid from a series of petite yeast strains by deoxyribonucleic acid-deoxyribonucleic acid hybridization. Biochemistry. 1974 Mar 12;13(6):1067–1075. doi: 10.1021/bi00703a002. [DOI] [PubMed] [Google Scholar]
  12. Hill B. T., Tsuboi A., Baserga R. Effect of 5-bromodeoxyuridine on chromatin transcription in confluent fibroblasts. Proc Natl Acad Sci U S A. 1974 Feb;71(2):455–459. doi: 10.1073/pnas.71.2.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Holmes D. S., Bonner J. Sequence composition of rat nuclear deoxyribonucleic acid and high molecular weight nuclear ribonucleic acid. Biochemistry. 1974 Feb 26;13(5):841–848. doi: 10.1021/bi00702a001. [DOI] [PubMed] [Google Scholar]
  14. Levitt D., Dorfman A. Control of chondrogenesis in limb-bud cell cultures by bromodeoxyuridine. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2201–2205. doi: 10.1073/pnas.70.8.2201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Levitt D., Dorfman A. The irreversible inhibition of differentiation of limb-bud mesenchyme by bromodeoxyuridine. Proc Natl Acad Sci U S A. 1972 May;69(5):1253–1257. doi: 10.1073/pnas.69.5.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lowy D. R., Rowe W. P., Teich N., Hartley J. W. Murine leukemia virus: high-frequency activation in vitro by 5-iododeoxyuridine and 5-bromodeoxyuridine. Science. 1971 Oct 8;174(4005):155–156. doi: 10.1126/science.174.4005.155. [DOI] [PubMed] [Google Scholar]
  17. Mccoll R. S., Aronson A. I. Transcription from unique and redundant DNA sequences in sea urchin embryos. Biochem Biophys Res Commun. 1974 Jan;56(1):47–51. doi: 10.1016/s0006-291x(74)80313-x. [DOI] [PubMed] [Google Scholar]
  18. Preisler H. S., Housman D., Scher W., Friend C. Effects of 5-bromo-2' -deoxyuridine on production of globin messenger RNA in dimethyl sulfoxide-stimulated Friend leukemia cells. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2956–2959. doi: 10.1073/pnas.70.10.2956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rosen J. M., Liarakos C. D., O'Malley B. W. Effect of estrogen on gene expression in the chick oviduct. I. Deoxyribonucleic acid--deoxyribonucleic acid renaturation studies. Biochemistry. 1973 Jul 17;12(15):2803–2809. doi: 10.1021/bi00739a005. [DOI] [PubMed] [Google Scholar]
  20. STUDIER F. W. SEDIMENTATION STUDIES OF THE SIZE AND SHAPE OF DNA. J Mol Biol. 1965 Feb;11:373–390. doi: 10.1016/s0022-2836(65)80064-x. [DOI] [PubMed] [Google Scholar]
  21. Schwartz S. A., Kirsten W. H. Differential response of rat tooth germs to a murine sarcoma virus in cell culture and syngeneic transplants. J Natl Cancer Inst. 1973 Oct;51(4):1163–1169. doi: 10.1093/jnci/51.4.1163. [DOI] [PubMed] [Google Scholar]
  22. Stellwagen R. H., Tomkins G. M. Preferential inhibition by 5-bromodeoxyuridine of the synthesis of tyrosine aminotransferase in hepatoma cell cultures. J Mol Biol. 1971 Feb 28;56(1):167–182. doi: 10.1016/0022-2836(71)90092-1. [DOI] [PubMed] [Google Scholar]
  23. Teich N., Lowy D. R., Hartley J. W., Rowe W. P. Studies of the mechanism of induction of infectious murine leukemia virus from AKR mouse embryo cell lines by 5-iododeoxyuridine and 5-bromodeoxyuridine. Virology. 1973 Jan;51(1):163–173. doi: 10.1016/0042-6822(73)90376-0. [DOI] [PubMed] [Google Scholar]
  24. Todaro G. J., Benveniste R. E., Lieber M. M., Livingston D. M. Infectious type C viruses released by normal cat embryo cells. Virology. 1973 Oct;55(2):506–515. doi: 10.1016/0042-6822(73)90192-x. [DOI] [PubMed] [Google Scholar]
  25. Varmus H. E., Bishop J. M., Vogt P. K. Appearance of virus-specific DNA in mammalian cells following transformation by Rous sarcoma virus. J Mol Biol. 1973 Mar 15;74(4):613–626. doi: 10.1016/0022-2836(73)90052-1. [DOI] [PubMed] [Google Scholar]
  26. Varmus H. E., Weiss R. A., Friis R. R., Levinson W., Bishop J. M. Detection of avian tumor virus-specific nucleotide sequences in avian cell DNAs (reassociation kinetics-RNA tumor viruses-gas antigen-Rous sarcoma virus, chick cells). Proc Natl Acad Sci U S A. 1972 Jan;69(1):20–24. doi: 10.1073/pnas.69.1.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Walther B. T., Pictet R. L., David J. D., Rutter W. J. On the mechanism of 5-bromodeoxyuridine inhibition of exocrine pancreas differentiation. J Biol Chem. 1974 Mar 25;249(6):1953–1964. [PubMed] [Google Scholar]
  28. Wetmur J. G., Davidson N. Kinetics of renaturation of DNA. J Mol Biol. 1968 Feb 14;31(3):349–370. doi: 10.1016/0022-2836(68)90414-2. [DOI] [PubMed] [Google Scholar]
  29. Wilt F. H., Anderson M. The action of 5-bromodeoxyuridine on differentiation. Dev Biol. 1972 Jun;28(2):443–447. doi: 10.1016/0012-1606(72)90026-7. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES