Cellular and Molecular Life Sciences

, Volume 66, Issue 19, pp 3219–3234

Functional interplay between Parp-1 and SirT1 in genome integrity and chromatin-based processes

  • Rosy El Ramy
  • Najat Magroun
  • Nadia Messadecq
  • Laurent R. Gauthier
  • François D. Boussin
  • Ullas Kolthur-Seetharam
  • Valérie Schreiber
  • Michael W. McBurney
  • Paolo Sassone-Corsi
  • Françoise Dantzer
Research Article

DOI: 10.1007/s00018-009-0105-4

Cite this article as:
El Ramy, R., Magroun, N., Messadecq, N. et al. Cell. Mol. Life Sci. (2009) 66: 3219. doi:10.1007/s00018-009-0105-4

Abstract

Poly(ADP-ribose) polymerase-1 (Parp-1) and the protein deacetylase SirT1 are two of the most effective NAD+-consuming enzymes in the cell with key functions in genome integrity and chromatin-based pathways. Here, we examined the in vivo crosstalk between both proteins. We observed that the double disruption of both genes in mice tends to increase late post-natal lethality before weaning consistent with important roles of both proteins in genome integrity during mouse development. We identified increased spontaneous telomeric abnormalities associated with decreased cell growth in the absence of either SirT1 or SirT1 and Parp-1 in mouse cells. In contrast, the additional disruption of Parp-1 rescued the abnormal pericentric heterochromatin, the nucleolar disorganization and the mitotic defects observed in SirT1-deficient cells. Together, these findings are in favor of key functions of both proteins in cellular response to DNA damage and in the modulation of histone modifications associated with constitutive heterochromatin integrity.

Keywords

Poly(ADP-ribosyl)ationAcetylationGenome integrityChromatin modificationsSirtuins

Supplementary material

18_2009_105_MOESM1_ESM.ppt (206 kb)
Sup.Fig.1. Retarded growth of Parp-1;SirT1 double knockout mice. Average weight curves of Parp-1+/+;SirT1+/+ (◆), Parp-1+/−;SirT1+/− (◊), Parp-1+/+;SirT1−/− (●), Parp-1+/−;SirT1−/− (▲) and Parp-1−/−;SirT1−/− (×) females (left panel) and males (right panel) ; n, number of mice included in the weigth curves. Parp-1−/−;SirT1−/− mice were 30-40% smaller compared to controls. For the female, this represents an exacerbation of the 20-30% reduction seen in Parp-1+/+; SirT1−/− mice (this study and (McBurney, M.W., Yang, X., Jardine, K., Hixon, M., Boekelheide, K., Webb, J.R., Lansdorp, P.M. and Lemieux, M. (2003). The mammalian SIR2alpha protein has a role in embryogenesis and gametogenesis. Mol Cell Biol 23, 38-54.)). (PPT 206 kb)
18_2009_105_MOESM2_ESM.ppt (4.4 mb)
Sup.Fig.2 (A) Normal patterns of H3K9me3 and HP1α in Parp-1-deficient cells. Representative immunofluorescence images for the comparative distribution of H3K9me3 (b,c) and HP1α (f,h) in Parp-1+/+;SirT1+/+ (a,b,e,f) and Parp-1−/−;SirT1+/+ (c,d,g,h) interphase cells. DNA and heterochromatic foci are counterstained with DAPI (a,c,e,g). Scale bars, 4,45 μm. (B) Dispersed distribution of H3K9me3 in Parp-1−/−;SirT1−/− cells reconstituted with Parp-1. Parp-1−/− ;SirT1−/− cells were transfected with either EGFP-Parp-1 or GST-Parp-1 and processed for immunofluorescence 36h later. a,d: DAPI-stained DNA. b: EGFP fluorescence. e: green labeled anti-GST antibody. c,f: red labeled anti anti-H3K9me3 antibody. Scale bars, 11 μm. (PPT 4483 kb)
18_2009_105_MOESM3_ESM.ppt (80 kb)
Sup.Fig.3 Western-blot analysis for the expression of SirT1 and β-actin in SirT1-transfected cells. Equivalent amounts of total protein extracts from mock-transfected (lanes 1,3) or pCruz-HA-SirT1 transfected (lanes 2,4) Parp-1+/+;SirT1+/+ (lanes 1-2) and Parp-1+/+;SirT1−/− (lanes 3-4) 3T3 cells were separated by SDS-PAGE and analyzed by Western blotting with the appropriate antibodies. (PPT 80 kb)

Copyright information

© Birkhäuser Verlag, Basel/Switzerland 2009

Authors and Affiliations

  • Rosy El Ramy
    • 1
  • Najat Magroun
    • 1
  • Nadia Messadecq
    • 2
  • Laurent R. Gauthier
    • 3
  • François D. Boussin
    • 3
  • Ullas Kolthur-Seetharam
    • 4
  • Valérie Schreiber
    • 1
  • Michael W. McBurney
    • 5
  • Paolo Sassone-Corsi
    • 6
  • Françoise Dantzer
    • 1
  1. 1.IREBS-FRE3211, ESBSIllkirchFrance
  2. 2.IGBMCIllkirchFrance
  3. 3.Laboratoire de RadiopathologieCEA, IRCM-INSERM U967Fontenay-aux-RosesFrance
  4. 4.Department of Biological SciencesTata Institute of Fundamental ResearchColabaIndia
  5. 5.Center for Cancer Therapeutics, Ottawa Health Research InstituteUniversity of OttawaOttawaCanada
  6. 6.Department of PharmacologyUniversity of California, GNRFIrvineUSA