Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1983;2(4):543–548. doi: 10.1002/j.1460-2075.1983.tb01460.x

Poly(ADP-ribose) polymerase auto-modification and interaction with DNA: electron microscopic visualization.

G de Murcia, J Jongstra-Bilen, M E Ittel, P Mandel, E Delain
PMCID: PMC555058  PMID: 6313345

Abstract

The interaction between purified calf thymus poly(ADP-ribose) polymerase and its activating co-purified DNA (sDNA) was investigated by electron microscopy. We have shown that the enzyme-DNA complex possesses a nucleosome-like structure. The enzyme-bound DNA (sDNA) was found to be enriched in single-stranded regions and branched structures, presumed to be replication forks. The auto-ribosylated polymerase as well as the branched poly(ADP-ribose) formed were visualized by dark field electron microscopy during the auto-ADP-ribosylation reaction and the possible mechanism of this phenomenon is discussed.

Full text

PDF
543

Images in this article

544Fig. 1.
on p.544
544Fig. 2.
on p.544
546Fig. 4.
on p.546
547Fig. 5.
on p.547

Selected References

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

  1. 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]
  2. Burzio L., Koide S. S. A functional role of polyADPR in DNA synthesis. Biochem Biophys Res Commun. 1970 Sep 10;40(5):1013–1020. doi: 10.1016/0006-291x(70)90894-6. [DOI] [PubMed] [Google Scholar]
  3. Caplan A. I., Rosenberg M. J. Interrelationship between poly (ADP-Rib) synthesis, intracellular NAD levels, and muscle or cartilage differentiation from mesodermal cells of embryonic chick limb. Proc Natl Acad Sci U S A. 1975 May;72(5):1852–1857. doi: 10.1073/pnas.72.5.1852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. De Murcia G., Wilhelm B., Wilhelm F. X., Daune M. P. Effect of tetramethylammonium ions on conformational changes of DNA in the premelting temperature range. Biophys Chem. 1978 Sep;8(4):377–383. doi: 10.1016/0301-4622(78)80019-2. [DOI] [PubMed] [Google Scholar]
  5. Dubochet J., Ducommun M., Zollinger M., Kellenberger E. A new preparation method for dark-field electron microscopy of biomacromolecules. J Ultrastruct Res. 1971 Apr;35(1):147–167. doi: 10.1016/s0022-5320(71)80148-x. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Ferro A. M., Olivera B. M. Poly(ADP-ribosylation) in vitro. Reaction parameters and enzyme mechanism. J Biol Chem. 1982 Jul 10;257(13):7808–7813. [PubMed] [Google Scholar]
  8. Hashida T., Ohgushi H., Yoshihara K. Highly effective DNA in stimulating poly-(ADP-ribose) polymerase reaction. Biochem Biophys Res Commun. 1979 May 14;88(1):305–311. doi: 10.1016/0006-291x(79)91730-3. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Hilz H. ADP-ribosylation of proteins--a multifunctional process. Hoppe Seylers Z Physiol Chem. 1981 Nov;362(11):1415–1425. [PubMed] [Google Scholar]
  11. Hilz H., Stone P. Poly(ADP-ribose) and ADP-ribosylation of proteins. Rev Physiol Biochem Pharmacol. 1976;76:1-58, 177. doi: 10.1007/BFb0027686. [DOI] [PubMed] [Google Scholar]
  12. Juarez-Salinas H., Levi V., Jacobson E. L., Jacobson M. K. Poly(ADP-ribose) has a branched structure in vivo. J Biol Chem. 1982 Jan 25;257(2):607–609. [PubMed] [Google Scholar]
  13. Jump D. B., Butt T. R., Smulson M. Nuclear protein modification and chromatin substructure. 3. Relationship between poly(adenosine diphosphate) ribosylation and different functional forms of chromatin. Biochemistry. 1979 Mar 20;18(6):983–990. doi: 10.1021/bi00573a008. [DOI] [PubMed] [Google Scholar]
  14. Kanai M., Miwa M., Kuchino Y., Sugimura T. Presence of branched portion in poly(adenosine diphosphate ribose) in vivo. J Biol Chem. 1982 Jun 10;257(11):6217–6223. [PubMed] [Google Scholar]
  15. Kawaichi M., Ueda K., Hayaishi O. Multiple autopoly(ADP-ribosyl)ation of rat liver poly(ADP-ribose) synthetase. Mode of modification and properties of automodified synthetase. J Biol Chem. 1981 Sep 25;256(18):9483–9489. [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Mandel P., Okazaki H., Niedergang C. Poly(adenosine diphosphate ribose). Prog Nucleic Acid Res Mol Biol. 1982;27:1–51. doi: 10.1016/s0079-6603(08)60596-6. [DOI] [PubMed] [Google Scholar]
  18. Mandel P., Okazaki H., Niedergang C. Purification and properties of calf thymus poly adenosine diphosphate ribose polymerase. FEBS Lett. 1977 Dec 15;84(2):331–336. doi: 10.1016/0014-5793(77)80719-9. [DOI] [PubMed] [Google Scholar]
  19. Maniatis T., Jeffrey A., van deSande H. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry. 1975 Aug 26;14(17):3787–3794. doi: 10.1021/bi00688a010. [DOI] [PubMed] [Google Scholar]
  20. Miwa M., Saikawa N., Yamaizumi Z., Nishimura S., Sugimura T. Structure of poly(adenosine diphosphate ribose): identification of 2'-[1''-ribosyl-2''-(or 3''-)(1'''-ribosyl)]adenosine-5',5'',5'''-tris(phosphate) as a branch linkage. Proc Natl Acad Sci U S A. 1979 Feb;76(2):595–599. doi: 10.1073/pnas.76.2.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Moyne G., Freeman R., Saragosti S., Yaniv M. A high-resolution electron microscopy study of nucleosomes from simian virus 40 chromatin. J Mol Biol. 1981 Jul 15;149(4):735–744. doi: 10.1016/0022-2836(81)90355-7. [DOI] [PubMed] [Google Scholar]
  22. Niedergang C., Okazaki H., Mandel P. Properties of purified calf thymus poly(adenosine diphosphate ribose) polymerase. Comparison of the DNA-independent and the DNA-dependent enzyme. Eur J Biochem. 1979 Dec;102(1):43–57. doi: 10.1111/j.1432-1033.1979.tb06261.x. [DOI] [PubMed] [Google Scholar]
  23. Nishikimi M., Ogasawara K., Kameshita I., Taniguchi T., Shizuta Y. Poly(ADP-ribose) synthetase. The DNA binding domain and the automodification domain. J Biol Chem. 1982 Jun 10;257(11):6102–6105. [PubMed] [Google Scholar]
  24. Ohgushi H., Yoshihara K., Kamiya T. Bovine thymus poly(adenosine diphosphate ribose) polymerase. Physical properties and binding to DNA. J Biol Chem. 1980 Jul 10;255(13):6205–6211. [PubMed] [Google Scholar]
  25. Okazaki H., Niedergang C., Mandel P. Adenosine diphosphate ribosylation of histone H1 by purified calf thymus polyadenosine diphosphate ribose polymerase. Biochimie. 1980;62(2-3):147–157. doi: 10.1016/s0300-9084(80)80190-8. [DOI] [PubMed] [Google Scholar]
  26. Okazaki H., Niedergang C., Mandel P. Purification and properties of calf thymus polyadenosine diphosphate ribose polymerase. FEBS Lett. 1976 Mar 1;62(3):255–258. doi: 10.1016/0014-5793(76)80069-5. [DOI] [PubMed] [Google Scholar]
  27. Petzold S. J., Booth B. A., Leimbach G. A., Berger N. A. Purification and properties of poly(ADP-ribose) polymerase from lamb thymus. Biochemistry. 1981 Dec 8;20(25):7075–7081. doi: 10.1021/bi00528a004. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. 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]
  30. Saragosti S., Moyne G., Yaniv M. Absence of nucleosomes in a fraction of SV40 chromatin between the origin of replication and the region coding for the late leader RNA. Cell. 1980 May;20(1):65–73. doi: 10.1016/0092-8674(80)90235-4. [DOI] [PubMed] [Google Scholar]
  31. Smulson M., Stark P., Gazzoli M., Roberts J. Release of template restriction for DNA synthesis by poly ADP(ribose) polymerase during the HeLa cell cycle. Exp Cell Res. 1975 Jan;90(1):175–182. doi: 10.1016/0014-4827(75)90371-7. [DOI] [PubMed] [Google Scholar]
  32. Whish W. J., Davies M. I., Shall S. Stimulation of poly(ADP-ribose) polymerase activity by the anti-tumour antibiotic, streptozotocin. Biochem Biophys Res Commun. 1975 Jul 22;65(2):722–730. doi: 10.1016/s0006-291x(75)80205-1. [DOI] [PubMed] [Google Scholar]
  33. Yoshihara K., Hashida T., Tanaka Y., Matsunami N., Yamaguchi A., Kamiya T. Mode of enzyme-bound poly(ADP-ribose) synthesis and histone modification by reconstituted poly(ADP-ribose) polymerase-DNA-cellulose complex. J Biol Chem. 1981 Apr 10;256(7):3471–3478. [PubMed] [Google Scholar]
  34. Zahradka P., Ebisuzaki K. A shuttle mechanism for DNA-protein interactions. The regulation of poly(ADP-ribose) polymerase. Eur J Biochem. 1982 Oct;127(3):579–585. [PubMed] [Google Scholar]
  35. de Murcia G., Das G. C., Erard M., Daune M. Superstructure and CD spectrum as probes of chromatin integrity. Nucleic Acids Res. 1978 Feb;5(2):523–535. doi: 10.1093/nar/5.2.523. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES