• A novel non-canonical PIP-box mediates PARG interaction with PCNA

    • Tanja Kaufmann
      Department of Biochemistry and Cell Biology, Centre for Molecular Biology, University of Vienna
    • Irina Grishkovskaya
      Department of Structural and Computational Biology, Centre for Molecular Biology, University of Vienna
    • Anton A. Polyansky
      Department of Structural and Computational Biology, Centre for Molecular Biology, University of Vienna
    • Sebastian Kostrhon
      Department of Biochemistry and Cell Biology, Centre for Molecular Biology, University of Vienna
    • Eva Kukolj
      Department of Biochemistry and Cell Biology, Centre for Molecular Biology, University of Vienna
    • Karin M. Olek
      Department of Biochemistry and Cell Biology, Centre for Molecular Biology, University of Vienna
    • Sebastien Herbert
      Department of Biochemistry and Cell Biology, Centre for Molecular Biology, University of Vienna
    • Etienne Beltzung
      Department of Biochemistry and Cell Biology, Centre for Molecular Biology, University of Vienna
    • Karl Mechtler
      Institute of Molecular Pathology, Vienna Biocenter (VBC)
    • Thomas Peterbauer
      Department of Biochemistry and Cell Biology, Centre for Molecular Biology, University of Vienna
    • Josef Gotzmann
      Department of Biochemistry and Cell Biology, Centre for Molecular Biology, University of Vienna
    • Lijuan Zhang
      VBCF-Advanced Microscopy, Vienna Biocenter (VBC)
    • Markus Hartl
      Large-Instrument Facility for Mass Spectrometry in Life Sciences, Faculty of Life Sciences, University of Vienna
    • Bojan Zagrovic
      Department of Structural and Computational Biology, Centre for Molecular Biology, University of Vienna
    • Kareem Elsayad
      VBCF-Advanced Microscopy, Vienna Biocenter (VBC)
    • Kristina Djinovic-Carugo
      Department of Structural and Computational Biology, Centre for Molecular Biology, University of Vienna
    • Dea Slade
      Department of Biochemistry and Cell Biology, Centre for Molecular Biology, University of Vienna
  • Poly(ADP-ribose) glycohydrolase (PARG) regulates cellular poly(ADP-ribose) (PAR) levels by rapidly cleaving glycosidic bonds between ADP-ribose units. PARG interacts with proliferating cell nuclear antigen (PCNA) and is strongly recruited to DNA damage sites in a PAR- and PCNA-dependent fashion. Here we identified PARG acetylation site K409 that is essential for its interaction with PCNA, its localization within replication foci and its recruitment to DNA damage sites. We found K409 to be part of a non-canonical PIP-box within the PARG disordered regulatory region. The previously identified putative N-terminal PIP-box does not bind PCNA directly but contributes to PARG localization within replication foci. X-ray structure and MD simulations reveal that the PARG non-canonical PIP-box binds PCNA in a manner similar to other canonical PIP-boxes and may represent a new type of PIP-box. While the binding of previously described PIP-boxes is based on hydrophobic interactions, PARG PIP-box binds PCNA via both stabilizing hydrophobic and fine-tuning electrostatic interactions. Our data explain the mechanism of PARG–PCNA interaction through a new PARG PIP-box that exhibits non-canonical sequence properties but a canonical mode of PCNA binding.

  • PDF

  • http://phaidra.univie.ac.at/o:907609

  • Article

  • Published Version

  • 2017

  • 45

  • 16

  • 9741-9759

  • Oxford University Press (OUP)

  • English

  • Open access

  • LS14-001 – Vienna Science and Technology Fund (WWTF)

  • W1258 – Austrian Science Fund (FWF)

  • 279408 – European Union (all programmes)

  • 0305-1048