Abstract
Poly(ADP-ribosyl)ation is a posttranslational modification of proteins in higher eukaryotes mediated by poly(ADP-ribose) polymerases (PARPs) that is involved in many physiological processes such as DNA repair, transcription, cell division, and cell death. Biochemical studies together with PARP-1- or PARP-2-deficient cellular and animal models have revealed the redundant but also complementary functions of the two enzymes in the surveillance and maintenance of genome integrity. Poly(ADP-ribose) is degraded by the endo- and exo-glycosidase activities of poly(ADP-ribose) glycohydrolase (PARG). In this chapter, biochemical and immunofluorescence methods are described for detecting and assaying PARPs and PARG.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kraus, W.L. and Lis, J.T. (2003) PARP goes transcription. Cell. 113, 677-683.
Amé, J.C., Spenlehauer, C., and de Murcia, G. (2004) The PARP superfamily. Bioessays. 26, 882-893.
Schreiber, V., Dantzer, F., Amé, J.C., and de Murcia, G. (2006) Poly(ADP-ribose): novel functions for an old molecule. Nat. Rev. Mol. Cell. Biol. 7, 517-528.
Ménissier de Murcia, J., Niedergang, C., Trucco, C., Ricoul, M., Dutrillaux, B., Mark, M., Olivier, F.J., Masson, M., Dierich, A., LeMeur, M., Walztinger, C., Chambon, P., and de Murcia, G. (1997) Requirement of poly(ADP-ribose)polymerase in recovery from DNA damage in mice and in cells. Proc. Natl. Acad. Sci. USA. 94, 7303-7307
Wang, Z.Q., Stingl, L., Morrison, C., Jantsch, M., Los, M., Schulze-Osthoff, K., and Wagner, E.F. (1997) PARP is important for genomic stability but dispensable in apoptosis. Genes Dev. 11, 2347-2358.
Masutani, M., Nozaki, T., Nishiyama, E., Shimokawa, T., Tachi, Y., Suzuki, H., Nakagama, H., Wakabayashi, K., and Sugimura, M. (1999) Function of poly(ADP-ribose) polymerase in response to DNA damage: gene-disruption study in mice. Mol. Cell. Biochem. 193, 149-152.
Schreiber, V., Amé, J.C., Dolle, P., Schultz, I., Rinaldi, B., Fraulob, V., Menissier-de Murcia, J., and de Murcia, G. (2002) Poly(ADP-ribose) polymerase-2 (PARP-2) is required for efficient base excision DNA repair in association with PARP-1 and XRCC1. J. Biol. Chem. 277, 23028-23036.
Ménissier de Murcia, J., Ricoul, M., Tartier, L., Niedergang, C., Huber, A., Dantzer, F., Schreiber, V., Amé, J.C., Dierich, A., LeMeur, M., Sabatier, L., Chambon, P., and de Murcia, G. (2003) Functional interaction between PARP-1 and PARP-2 in chromosome stability and embryonic development in mouse. EMBO J. 22, 2255-2263.
Amé, J.C., Jacobson, E.L., and Jacobson, M.K. (1999) Molecular heterogeneity and regulation of poly(ADP-ribose) glycohydrolase. Mol. Cell. Biochem. 193, 75-81.
Meyer-Ficca, M.L., Meyer, R.G., Coyle, D.L., Jacobson, E.L., and Jacobson, M.K. (2004) Human poly(ADP-ribose) glycohydrolase is expressed in alternative splice variants yielding isoforms that localize to different cell compartments. Exp. Cell. Res. 297, 521-532.
Andrabi, S.A., Kim, N.S., Yu, S.W., Wang, H., Koh, D.W., Sasaki, M., Klaus, J.A., Otsuka, T., Zhang, Z., Koehler, R.C., Hurn, P.D., Poirier, G.G., Dawson, V.L., and Dawson, T.M. (2006) Poly(ADP-ribose) (PAR) polymer is a death signal. Proc. Natl. Acad. Sci. USA. 103, 18308-18313.
Gagné, J.P., Hendzel, M.J., Droit, A., and Poirier, G.G. (2006) The expanding role of poly(ADP-ribose) metabolism: current challenges and new perspectives. Curr. Opin. Cell. Biol. 18, 145-151.
Lin, W., Amé, J.C., Aboul-Ela, N., Jacobson, E.L., and Jacobson, M.K. (1997) Isolation and characterization of the cDNA encoding bovine poly(ADP-ribose) glycohydrolase. J. Biol. Chem. 272, 11895-11901.
Méder, V.S., Boeglin, M., de Murcia, G., and Schreiber, V. (2005) PARP-1 and PARP-2 interact with nucleophosmin/B23 and accumulate in transcriptionally active nucleoli. J. Cell Sci. 118, 211-222.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science + Business Media, LLC
About this protocol
Cite this protocol
Amé, JC., Hakmé, A., Quenet, D., Fouquerel, E., Dantzer, F., Schreiber, V. (2008). Detection of the Nuclear Poly(ADP-ribose)-Metabolizing Enzymes and Activities in Response to DNA Damage. In: Hancock, R. (eds) The Nucleus. Methods in Molecular Biology, vol 464. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-461-6_15
Download citation
DOI: https://doi.org/10.1007/978-1-60327-461-6_15
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-460-9
Online ISBN: 978-1-60327-461-6
eBook Packages: Springer Protocols