Abstract
Reversible post-translational modification by poly(ADP-ribose) (PAR) regulates chromatin structure, DNA repair and cell fate in response to genotoxic stress. PAR glycohydrolase (PARG) removes PAR chains from poly ADP-ribosylated proteins to restore protein function and release oligo(ADP-ribose) chains to signal damage. Here we report crystal structures of mammalian PARG and its complex with a substrate mimic that reveal an open substrate-binding site and a unique 'tyrosine clasp' enabling endoglycosidic cleavage of branched PAR chains.
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Acknowledgements
We thank W.L. Kraus at University of Texas Southwestern Medical Center for providing a pET28a-ratPARG(1–972) plasmid. This work was supported in part by grants from the US National Institutes of Health (including 5R01 GM052504 to T.E.) and the Structural Cell Biology of DNA Repair Program (P01 CA92584 to T.E. and J.A.T.). X-ray diffraction and scattering technologies and their applications to the determination of macromolecular structures and conformations at the SIBYLS beamline at the Advanced Light Source, Lawrence Berkeley National Laboratory, are supported in part by the US Department of Energy program Integrated Diffraction Analysis Technologies under Contract Number DE-AC02-05CH11231 with the US Department of Energy.
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I.-K.K. designed experiments, crystallized the proteins, determined the crystal structures, analyzed results and wrote the manuscript. J.R.K. designed experiments, analyzed results and wrote the manuscript. C.M.W.H. performed the computational simulations. R.A.S. purified and crystallized the proteins. S.C. and J.A.T. helped with X-ray data collection and writing the manuscript. T.E. analyzed results, wrote the manuscript and provided financial support.
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Kim, IK., Kiefer, J., Ho, C. et al. Structure of mammalian poly(ADP-ribose) glycohydrolase reveals a flexible tyrosine clasp as a substrate-binding element. Nat Struct Mol Biol 19, 653–656 (2012). https://doi.org/10.1038/nsmb.2305
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DOI: https://doi.org/10.1038/nsmb.2305
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