This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kumari, S. R. Articles by Alvarez-Gonzalez, R. Search for Related Content PubMed PubMed Citation Articles by Kumari, S. R. Articles by Alvarez-Gonzalez, R.

Functional Interactions of p53 with Poly(ADP-ribose) Polymerase (PARP) during Apoptosis following DNA Damage: Covalent Poly(ADP-ribosyl)ation of p53 by Exogenous PARP and Noncovalent Binding of p53 to the M_r 85,000 Proteolytic Fragment¹

Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, Texas 76107

We have examined the domain-specific interactions between p53 and poly(ADP-ribose)polymerase (PARP) (E.C. 2.4.2.30) in apoptotic HeLa cells. Apoptosis was induced by exposing cells to 50 µM N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) for increasing lengths of time and was confirmed by: (a) oligonucleosomal fragmentation of chromatin; (b) increase in p53 levels; and (c) degradation of PARP into the characteristic M_r 85,000 (COOH-terminal catalytic domain) and M_r 29,000 (DNA-binding domain) peptide fragments. We also immunodetected p53 in immunoprecipitates obtained with a PARP-specific antibody. However, intact PARP coimmunoprecipitated with a p53-specific antibody during the initial 30 min of MNNG treatment. After 60 min, only the COOH-terminal fragment coimmunoprecipitated with p53, indicating that PARP noncovalently binds p53 via its M_r 85,000 catalytic domain. Therefore, we next examined p53 as a covalent target for poly(ADP-ribosyl)ation. Although p53 was not endogenously poly(ADP-ribosyl)ated in situ, incubation of cell extracts with full-length PARP from calf thymus and [³²P]ßNAD⁺ resulted in its time-dependent poly(ADP-ribosyl)ation. In summary, our results are consistent with the conclusion that PARP and p53 are activated with nonoverlapping kinetics during apoptosis.

¹ This project was supported by Grant 9678-014 from the Texas Advanced Research Program and Grant GM45451 from the NIH.

² To whom requests for reprints should be addressed, at Department of Molecular Biology and Immunology, University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107-2699. Phone: (817) 735-2117; Fax: (817) 735-2118; E-mail: ralvarez@hsc.unt.edu.

Received 7/14/98. Accepted 9/24/98.

This article has been cited by other articles:

				T. Di Palma, T. de Cristofaro, C. D'Ambrosio, D. Del Prete, A. Scaloni, and M. Zannini Poly(ADP-ribose) polymerase 1 binds to Pax8 and inhibits its transcriptional activity J. Mol. Endocrinol., November 1, 2008; 41(5): 379 - 388. [Abstract] [Full Text] [PDF]

				R. V. Warke, K. J. Martin, K. Giaya, S. K. Shaw, A. L. Rothman, and I. Bosch TRAIL Is a Novel Antiviral Protein against Dengue Virus J. Virol., January 1, 2008; 82(1): 555 - 564. [Abstract] [Full Text] [PDF]

				R. Alvarez-Gonzalez Genomic Maintenance: The p53 Poly(ADP-ribosyl)ation Connection Sci. Signal., December 4, 2007; 2007(415): pe68 - pe68. [Abstract] [Full Text] [PDF]

				S. Beneke and A. Burkle Poly(ADP-ribosyl)ation in mammalian ageing Nucleic Acids Res., December 3, 2007; 35(22): 7456 - 7465. [Abstract] [Full Text] [PDF]

				C. Baumann, G. S. Boehden, A. Burkle, and L. Wiesmuller Poly(ADP-RIBOSE) polymerase-1 (Parp-1) antagonizes topoisomerase I-dependent recombination stimulation by P53 Nucleic Acids Res., February 9, 2006; 34(3): 1036 - 1049. [Abstract] [Full Text] [PDF]

				S. Beneke and A. Burkle Poly(ADP-Ribosyl)ation, PARP, and Aging Sci. Aging Knowl. Environ., December 8, 2004; 2004(49): re9 - re9. [Abstract] [Full Text] [PDF]

				S. Susse, C.-J. Scholz, A. Burkle, and L. Wiesmuller Poly(ADP-ribose) polymerase (PARP-1) and p53 independently function in regulating double-strand break repair in primate cells Nucleic Acids Res., February 2, 2004; 32(2): 669 - 680. [Abstract] [Full Text] [PDF]

				A. W. Oliver, J.-C. Ame, S. M. Roe, V. Good, G. de Murcia, and L. H. Pearl Crystal structure of the catalytic fragment of murine poly(ADP-ribose) polymerase-2 Nucleic Acids Res., January 22, 2004; 32(2): 456 - 464. [Abstract] [Full Text] [PDF]

				S. Ishizuka, K. Martin, C. Booth, C. S. Potten, G. de Murcia, A. Burkle, and T. B. L. Kirkwood Poly(ADP-ribose) polymerase-1 is a survival factor for radiation-exposed intestinal epithelial stem cells in vivo Nucleic Acids Res., November 1, 2003; 31(21): 6198 - 6205. [Abstract] [Full Text] [PDF]

				M. Hasmann and I. Schemainda FK866, a Highly Specific Noncompetitive Inhibitor of Nicotinamide Phosphoribosyltransferase, Represents a Novel Mechanism for Induction of Tumor Cell Apoptosis Cancer Res., November 1, 2003; 63(21): 7436 - 7442. [Abstract] [Full Text] [PDF]

				S. Wieler, J.-P. Gagne, H. Vaziri, G. G. Poirier, and S. Benchimol Poly(ADP-ribose) Polymerase-1 Is a Positive Regulator of the p53-mediated G1 Arrest Response following Ionizing Radiation J. Biol. Chem., May 23, 2003; 278(21): 18914 - 18921. [Abstract] [Full Text] [PDF]

				L. Virag and C. Szabo The Therapeutic Potential of Poly(ADP-Ribose) Polymerase Inhibitors Pharmacol. Rev., September 1, 2002; 54(3): 375 - 429. [Abstract] [Full Text] [PDF]

				W.-J. Chang and R. Alvarez-Gonzalez The Sequence-specific DNA Binding of NF-kappa B Is Reversibly Regulated by the Automodification Reaction of Poly (ADP-ribose) Polymerase 1 J. Biol. Chem., December 7, 2001; 276(50): 47664 - 47670. [Abstract] [Full Text] [PDF]

				M. N. Cervellera and A. Sala Poly(ADP-ribose) Polymerase Is a B-MYB Coactivator J. Biol. Chem., March 31, 2000; 275(14): 10692 - 10696. [Abstract] [Full Text] [PDF]

				R. Alvarez-Gonzalez, H. Spring, M. Muller, and A. Burkle Selective Loss of Poly(ADP-ribose) and the 85-kDa Fragment of Poly(ADP-ribose) Polymerase in Nucleoli during Alkylation-induced Apoptosis of HeLa Cells J. Biol. Chem., November 5, 1999; 274(45): 32122 - 32126. [Abstract] [Full Text] [PDF]

				H. M. Smith and A. J. Grosovsky PolyADP-ribose-mediated regulation of p53 complexed with topoisomerase I following ionizing radiation Carcinogenesis, August 1, 1999; 20(8): 1439 - 1444. [Abstract] [Full Text] [PDF]

				C. Guo, S. Yu, A. T. Davis, H. Wang, J. E. Green, and K. Ahmed A Potential Role of Nuclear Matrix-associated Protein Kinase CK2 in Protection against Drug-induced Apoptosis in Cancer Cells J. Biol. Chem., February 16, 2001; 276(8): 5992 - 5999. [Abstract] [Full Text] [PDF]

				A. H. Boulares, A. J. Zoltoski, A. Yakovlev, M. Xu, and M. E. Smulson Roles of DNA Fragmentation Factor and Poly(ADP-ribose) Polymerase in an Amplification Phase of Tumor Necrosis Factor-induced Apoptosis J. Biol. Chem., October 5, 2001; 276(41): 38185 - 38192. [Abstract] [Full Text] [PDF]

				H. Mendoza-Alvarez and R. Alvarez-Gonzalez Regulation of p53 Sequence-specific DNA-binding by Covalent Poly(ADP-ribosyl)ation J. Biol. Chem., September 21, 2001; 276(39): 36425 - 36430. [Abstract] [Full Text] [PDF]