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Role of PARP-1 and PARP-2 in the expression of apoptosis-regulating genes in HeLa cells

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Abstract

Poly (ADP-ribose) polymerase-1 (PARP-1) is a DNA-binding enzyme involved in DNA damage processing, apoptosis, and genetic stability. Many lines of evidence suggest that PARP-1 is implicated in transcriptional regulation of various genes through the modulation of chromatin structure or through direct interaction with transcription factors and/or transcription factor-binding sites. In the present study, we applied TaqMan Low-Density Array analyses to investigate the expression of genes involved in apoptotic cell death induced by an alkylating agent. Using RNA interference, we determined the roles of PARP-1 and PARP-2 in transcriptional regulation during apoptosis in HeLa cells. Of the 93 genes monitored, 33 differentially expressed genes were identified after induction of apoptosis. Whereas the down-regulation of PARP-1 and PARP-2 had no impact on gene expression per se, we observed that Bcl10, c-Rel, and tumor necrosis factor-related apoptosis-inducing ligand receptor-1 and -2 are differentially expressed after induction of apoptosis in a PARP-1-dependent manner. These findings suggest that PARP-1—but not PARP-2—is required for proper expression of major genes involved in regulation of apoptosis.

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Abbreviations

ACTB:

beta-actin

Bcl10:

B-cell lymphoma 10

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

fetal bovine serum

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

MNNG:

N-methyl-N′-nitro-N′-nitrosoguanidine

PAR:

poly(ADP-ribose)

PARP-1:

PAR polymerase 1

PARP-2:

PAR polymerase 2

RNAi:

RNA interference

siRNA:

small interfering RNA

TLDA:

TaqMan® Low-Density Array

TRAIL-R1 and -R2:

TNF-related apoptosis-inducing ligand receptor-1 and -2

References

  • Abbadie C, Kabrun N, Bouali F, Smardova J, Stehelin D, Vandenbunder B, et al. High levels of c-rel expression are associated with programmed cell death in the developing avian embryo and in bone marrow cells in vitro. Cell 1993;75:899–912.

    Article  PubMed  CAS  Google Scholar 

  • Akiyama T, Takasawa S, Nata K, Kobayashi S, Abe M, Shervani NJ, et al. Activation of Reg gene, a gene for insulin-producing beta-cell regeneration: poly(ADP-ribose) polymerase binds Reg promoter and regulates the transcription by autopoly(ADP-ribosyl)ation. Proc Natl Acad Sci U S A 2001;98:48–53.

    Article  PubMed  CAS  Google Scholar 

  • Althaus FR. Poly ADP-ribosylation: a histone shuttle mechanism in DNA excision repair. J Cell Sci. 1992;102(Pt 4):663–70.

    PubMed  CAS  Google Scholar 

  • Ame JC, Rolli V, Schreiber V, Niedergang C, Apiou F, Decker P, et al. PARP-2, A novel mammalian DNA damage-dependent poly(ADP-ribose) polymerase. J Biol Chem. 1999;274:17860–8.

    Article  PubMed  CAS  Google Scholar 

  • Attar RM, Macdonald-Bravo H, Raventos-Suarez C, Durham SK, Bravo R. Expression of constitutively active IkappaB beta in T cells of transgenic mice: persistent NF-kappaB activity is required for T-cell immune responses. Mol Cell Biol. 1998;18:477–87.

    PubMed  CAS  Google Scholar 

  • Beg AA, Baltimore D. An essential role for NF-kappaB in preventing TNF-alpha-induced cell death. Science 1996;274:782–4.

    Article  PubMed  CAS  Google Scholar 

  • Butler AJ, Ordahl CP. Poly(ADP-ribose) polymerase binds with transcription enhancer factor 1 to MCAT1 elements to regulate muscle-specific transcription. Mol Cell Biol. 1999;19:296–306.

    PubMed  CAS  Google Scholar 

  • Cervellera MN, Sala A. Poly(ADP-ribose) polymerase is a B-MYB coactivator. J Biol Chem. 2000;275:10692–6.

    Article  PubMed  CAS  Google Scholar 

  • Chen X, Kandasamy K, Srivastava RK. Differential roles of RelA (p65) and c-Rel subunits of nuclear factor kappa B in tumor necrosis factor-related apoptosis-inducing ligand signaling. Cancer Res. 2003;63:1059–66.

    PubMed  CAS  Google Scholar 

  • Choi HS, Hwang CK, Kim CS, Song KY, Law PY, Loh HH, et al. Transcriptional regulation of mouse mu opioid receptor gene in neuronal cells by Poly(ADP-ribose) polymerase-1. J Cell Mol Med. 2008 (in press).

  • Cohausz O, Blenn C, Malanga M, Althaus FR. The roles of poly(ADP-ribose)-metabolizing enzymes in alkylation-induced cell death. Cell Mol Life Sci. 2008;65(4):644–55.

    Article  PubMed  CAS  Google Scholar 

  • D’Amours D, Desnoyers S, D’Silva I, Poirier GG. Poly(ADP-ribosyl)ation reactions in the regulation of nuclear functions. Biochem J. 1999;342(Pt 2):249–68.

    Article  PubMed  Google Scholar 

  • Falschlehner C, Emmerich CH, Gerlach B, Walczak H. TRAIL signalling: decisions between life and death. Int J Biochem Cell Biol. 2007;39:1462–75.

    Article  PubMed  CAS  Google Scholar 

  • Golstein P. Cell death: TRAIL and its receptors. Curr Biol. 1997;7:R750–3.

    Article  PubMed  CAS  Google Scholar 

  • Haince JF, Rouleau M, Poirier GG. Transcription. Gene expression needs a break to unwind before carrying on. Science 2006;312:1752–3.

    Article  PubMed  CAS  Google Scholar 

  • Hassa PO, Hottiger MO. The functional role of poly(ADP-ribose)polymerase 1 as novel coactivator of NF-kappaB in inflammatory disorders. Cell Mol Life Sci. 2002;59:1534–53.

    Article  PubMed  CAS  Google Scholar 

  • Hassa PO, Buerki C, Lombardi C, Imhof R, Hottiger MO. Transcriptional coactivation of nuclear factor-kappaB-dependent gene expression by p300 is regulated by poly(ADP)-ribose polymerase-1. J Biol Chem. 2003;278:45145–53.

    Article  PubMed  CAS  Google Scholar 

  • Hassa PO, Haenni SS, Elser M, Hottiger MO. Nuclear ADP-ribosylation reactions in mammalian cells: where are we today and where are we going? Microbiol Mol Biol Rev. 2006;70:789–829.

    Article  PubMed  CAS  Google Scholar 

  • Hosokawa Y. Anti-apoptotic action of API2-MALT1 fusion protein involved in t(11;18)(q21;q21) MALT lymphoma. Apoptosis 2005;10:25–34.

    Article  PubMed  CAS  Google Scholar 

  • Ju BG, Solum D, Song EJ, Lee KJ, Rose DW, Glass CK, et al. Activating the PARP-1 sensor component of the groucho/TLE1 corepressor complex mediates a CaMKinase IIdelta-dependent neurogenic gene activation pathway. Cell 2004;119:815–29.

    Article  PubMed  CAS  Google Scholar 

  • Ju BG, Lunyak VV, Perissi V, Garcia-Bassets I, Rose DW, Glass CK, et al. A topoisomerase IIbeta-mediated dsDNA break required for regulated transcription. Science 2006;312:1798–802.

    Article  PubMed  CAS  Google Scholar 

  • Kannan P, Yu Y, Wankhade S, Tainsky MA. PolyADP-ribose polymerase is a coactivator for AP-2-mediated transcriptional activation. Nucleic Acids Res. 1999;27:866–74.

    Article  PubMed  CAS  Google Scholar 

  • Karin M, Lin A. NF-kappaB at the crossroads of life and death. Nat Immunol. 2002;3:221–7.

    Article  PubMed  CAS  Google Scholar 

  • Kim MY, Mauro S, Gevry N, Lis JT, Kraus WL. NAD+-dependent modulation of chromatin structure and transcription by nucleosome binding properties of PARP-1. Cell 2004;119:803–14.

    Article  PubMed  CAS  Google Scholar 

  • Koseki T, Inohara N, Chen S, Carrio R, Merino J, Hottiger MO, et al. CIPER, a novel NF kappaB-activating protein containing a caspase recruitment domain with homology to Herpesvirus-2 protein E10. J Biol Chem. 1999;274:9955–61.

    Article  PubMed  CAS  Google Scholar 

  • Kraus WL, Lis JT. PARP goes transcription. Cell 2003;113:677–83.

    Article  PubMed  CAS  Google Scholar 

  • Krishnakumar R, Gamble MJ, Frizzell KM, Berrocal JG, Kininis M, Kraus WL. Reciprocal binding of PARP-1 and histone H1 at promoters specifies transcriptional outcomes. Science 2008;319:819–21.

    Article  PubMed  CAS  Google Scholar 

  • Lipton SA. Janus faces of NF-kappa B: neurodestruction versus neuroprotection. Nat Med. 1997;3:20–2.

    Article  PubMed  CAS  Google Scholar 

  • Maeda Y, Hunter TC, Loudy DE, Dave V, Schreiber V, Whitsett JA. PARP-2 interacts with TTF-1 and regulates expression of surfactant protein-B. J Biol Chem. 2006;281:9600–6.

    Article  PubMed  CAS  Google Scholar 

  • Miyamoto T, Kakizawa T, Hashizume K. Inhibition of nuclear receptor signalling by poly(ADP-ribose) polymerase. Mol Cell Biol. 1999;19:2644–9.

    PubMed  CAS  Google Scholar 

  • Nie J, Sakamoto S, Song D, Qu Z, Ota K, Taniguchi T. Interaction of Oct-1 and automodification domain of poly(ADP-ribose) synthetase. FEBS Lett 1998;424:27–32.

    Article  PubMed  CAS  Google Scholar 

  • Nirodi C, NagDas S, Gygi SP, Olson G, Aebersold R, Richmond A. A role for poly(ADP-ribose) polymerase in the transcriptional regulation of the melanoma growth stimulatory activity (CXCL1) gene expression. J Biol Chem. 2001;276:9366–74.

    Article  PubMed  CAS  Google Scholar 

  • Oei SL, Griesenbeck J, Schweiger M, Babich V, Kropotov A, Tomilin N. Interaction of the transcription factor YY1 with human poly(ADP-ribosyl) transferase. Biochem Biophys Res Commun. 1997;240:108–11.

    Article  PubMed  CAS  Google Scholar 

  • Oliver FJ, Menissier-de Murcia J, Nacci C, Decker P, Andriantsitohaina R, Muller S, et al. Resistance to endotoxic shock as a consequence of defective NF-kappaB activation in poly (ADP-ribose) polymerase-1 deficient mice. Embo J. 1999;18:4446–54.

    Article  PubMed  CAS  Google Scholar 

  • Ravi R, Bedi GC, Engstrom LW, Zeng Q, Mookerjee B, Gelinas C, et al. Regulation of death receptor expression and TRAIL/Apo2L-induced apoptosis by NF-kappaB. Nat Cell Biol. 2001;3:409–16.

    Article  PubMed  CAS  Google Scholar 

  • Realini CA, Althaus FR. Histone shuttling by poly(ADP-ribosylation). J Biol Chem. 1992;267:18858–65.

    PubMed  CAS  Google Scholar 

  • Ryan KM, Ernst MK, Rice NR, Vousden KH. Role of NF-kappaB in p53-mediated programmed cell death. Nature 2000;404:892–7.

    Article  PubMed  CAS  Google Scholar 

  • Schreiber V, Dantzer F, Ame JC, de Murcia G. Poly(ADP-ribose): novel functions for an old molecule. Nat Rev Mol Cell Biol. 2006;7:517–28.

    Article  PubMed  CAS  Google Scholar 

  • Soldatenkov VA, Chasovskikh S, Potaman VN, Trofimova I, Smulson ME, Dritschilo A. Transcriptional repression by binding of poly(ADP-ribose) polymerase to promoter sequences. J Biol Chem. 2002;277:665–70.

    Article  PubMed  CAS  Google Scholar 

  • Thome M, Martinon F, Hofmann K, Rubio V, Steiner V, Schneider P, et al. Equine herpesvirus-2 E10 gene product, but not its cellular homologue, activates NF-kappaB transcription factor and c-Jun N-terminal kinase. J Biol Chem. 1999;274:9962–8.

    Article  PubMed  CAS  Google Scholar 

  • Tulin A, Spradling A. Chromatin loosening by poly(ADP)-ribose polymerase (PARP) at Drosophila puff loci. Science 2003;299:560–2.

    Article  PubMed  CAS  Google Scholar 

  • Van Antwerp DJ, Martin SJ, Kafri T, Green DR, Verma IM. Suppression of TNF-alpha-induced apoptosis by NF-kappaB. Science 1996;274:787–9.

    Article  PubMed  Google Scholar 

  • Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002;3:1–12.

    Article  Google Scholar 

  • Wesierska-Gadek J, Wojciechowski J, Schmid G. Phosphorylation regulates the interaction and complex formation between wt p53 protein and PARP-1. J Cell Biochem. 2003;89:1260–84.

    Article  PubMed  CAS  Google Scholar 

  • Willis TG, Jadayel DM, Du MQ, Peng H, Perry AR, Abdul-Rauf M, et al. Bcl10 is involved in t(1;14)(p22;q32) of MALT B cell lymphoma and mutated in multiple tumor types. Cell 1999;96:35–45.

    Article  PubMed  CAS  Google Scholar 

  • Yan M, Lee J, Schilbach S, Goddard A, Dixit V. mE10, a novel caspase recruitment domain-containing proapoptotic molecule. J Biol Chem. 1999;274:10287–92.

    Article  PubMed  CAS  Google Scholar 

  • Zhang Z, Hildebrandt EF, Simbulan-Rosenthal CM, Anderson MG. Sequence-specific binding of poly(ADP-ribose) polymerase-1 to the human T cell leukemia virus type-I tax responsive element. Virology 2002;296:107–16.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

We thank Dr. Andreas Bergthaler and the Institute of Experimental Immunology, University Hospital Zurich, for providing access to an ABI PRISM 7900HT Sequence Detection System, and Prof. Dr. Markus Doherr, for assistance and helpful discussions with the statistical analyses. This work was supported by a grant from the Swiss National Foundation for scientific research awarded to F.R.A.

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  1. Institute of Pharmacology and Toxicology, University of Zurich-Vetsuisse, Winterthurerstrasse 260, CH-8057, Zurich, Switzerland

    Odile Cohausz & Felix R. Althaus

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  1. Odile Cohausz
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  2. Felix R. Althaus
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Correspondence to Felix R. Althaus.

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Cohausz, O., Althaus, F.R. Role of PARP-1 and PARP-2 in the expression of apoptosis-regulating genes in HeLa cells. Cell Biol Toxicol 25, 379–391 (2009). https://doi.org/10.1007/s10565-008-9092-8

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