Skip to main content
PMC 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
. 1985 Jan;82(1):104–108. doi: 10.1073/pnas.82.1.104

Accumulation of 10-kilobase DNA replication intermediates in cells treated with 3-aminobenzamide.

U Lönn, S Lönn
PMCID: PMC396980  PMID: 2982139

Abstract

During eukaryotic DNA synthesis there is formation of, in addition to Okazaki fragments, discrete 10-kilobase (kb) DNA replication intermediates. We have investigated the ligation of 10-kb DNA replication intermediates to high molecular weight DNA, using the drug 3-aminobenzamide, an inhibitor of poly(ADP-ribose) synthetase. In human melanoma cells treated with this inhibitor, there is an accumulation of 10-kb DNA. In contrast, in cells treated with aphidicolin, which inhibits DNA polymerase alpha, there is continued ligation of 10-kb DNA to high molecular weight DNA. Furthermore, using sequential treatment with aphidicolin and 3-aminobenzamide, one can observe the conversion of radiolabeled Okazaki fragments into 10-kb intermediates. The 10-kb DNA pieces are, however, not ligated to high molecular weight DNA in the presence of 3-aminobenzamide. Our results imply that functioning poly(ADP-ribose) synthetase is necessary for the ligation process.

Full text

PDF
108

Selected References

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

  1. Aubin R. J., Fréchette A., de Murcia G., Mandel P., Lord A., Grondin G., Poirier G. G. Correlation between endogenous nucleosomal hyper(ADP-ribosyl)ation of histone H1 and the induction of chromatin relaxation. EMBO J. 1983;2(10):1685–1693. doi: 10.1002/j.1460-2075.1983.tb01643.x. [DOI] [PMC free article] [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. 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]
  4. Durkacz B. W., Omidiji O., Gray D. A., Shall S. (ADP-ribose)n participates in DNA excision repair. Nature. 1980 Feb 7;283(5747):593–596. doi: 10.1038/283593a0. [DOI] [PubMed] [Google Scholar]
  5. Giri C. P., West M. H., Ramirez M. L., Smulson M. Nuclear protein modification and chromatin substructure. 2. Internucleosomal localization of poly(adenosine diphosphate-ribose) polymerase. Biochemistry. 1978 Aug 22;17(17):3501–3504. doi: 10.1021/bi00610a012. [DOI] [PubMed] [Google Scholar]
  6. Hayaishi O., Ueda K. Poly(ADP-ribose) and ADP-ribosylation of proteins. Annu Rev Biochem. 1977;46:95–116. doi: 10.1146/annurev.bi.46.070177.000523. [DOI] [PubMed] [Google Scholar]
  7. Huberman J. A. New views of the biochemistry of eucaryotic DNA replication revealed by aphidicolin, an unusual inhibitor of DNA polymerase alpha. Cell. 1981 Mar;23(3):647–648. doi: 10.1016/0092-8674(81)90426-8. [DOI] [PubMed] [Google Scholar]
  8. James M. R., Lehmann A. R. Role of poly(adenosine diphosphate ribose) in deoxyribonucleic acid repair in human fibroblasts. Biochemistry. 1982 Aug 17;21(17):4007–4013. doi: 10.1021/bi00260a016. [DOI] [PubMed] [Google Scholar]
  9. Kidwell W. R., Mage M. G. Changes in poly(adenosine diphosphate-ribose) and poly(adenosine diphosphate-ribose) polymerase in synchronous HeLa cells. Biochemistry. 1976 Mar 23;15(6):1213–1217. doi: 10.1021/bi00651a006. [DOI] [PubMed] [Google Scholar]
  10. Lönn U. Delayed flow-through cytoplasm of newly synthesized Balbiani ring 75S RNA. Cell. 1978 Apr;13(4):727–733. doi: 10.1016/0092-8674(78)90222-2. [DOI] [PubMed] [Google Scholar]
  11. Lönn U. Detection of a 10 kb DNA replication intermediate in human melanoma cells. Chromosoma. 1982;84(5):663–673. doi: 10.1007/BF00286332. [DOI] [PubMed] [Google Scholar]
  12. Lönn U., Lönn S. Aphidicolin inhibits the synthesis and joining of short DNA fragments but not the union of 10-kilobase DNA replication intermediates. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3996–3999. doi: 10.1073/pnas.80.13.3996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Morgan W. F., Cleaver J. E. Effect of 3-aminobenzamide on the rate of ligation during repair of alkylated DNA in human fibroblasts. Cancer Res. 1983 Jul;43(7):3104–3107. [PubMed] [Google Scholar]
  14. Mullins D. W., Jr, Giri C. P., Smulson M. Poly(adenosine diphosphate-ribose) polymerase: the distribution of a chromosome-associated enzyme within the chromatin substructure. Biochemistry. 1977 Feb 8;16(3):506–513. doi: 10.1021/bi00622a026. [DOI] [PubMed] [Google Scholar]
  15. Ohashi Y., Ueda K., Kawaichi M., Hayaishi O. Activation of DNA ligase by poly(ADP-ribose) in chromatin. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3604–3607. doi: 10.1073/pnas.80.12.3604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Poirier G. G., de Murcia G., Jongstra-Bilen J., Niedergang C., Mandel P. Poly(ADP-ribosyl)ation of polynucleosomes causes relaxation of chromatin structure. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3423–3427. doi: 10.1073/pnas.79.11.3423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Purnell M. R., Stone P. R., Whish W. J. ADP-ribosylation of nuclear proteins. Biochem Soc Trans. 1980 Apr;8(2):215–227. doi: 10.1042/bst0080215. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Ueda K., Omachi A., Kawaichi M., Hayaishi O. Natural occurrence of poly(ADP-ribosyl) histones in rat liver. Proc Natl Acad Sci U S A. 1975 Jan;72(1):205–209. doi: 10.1073/pnas.72.1.205. [DOI] [PMC free article] [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