Abstract
Upon binding to DNA breaks, poly(ADP-ribose) polymerase 1 (PARP1) ADP-ribosylates itself and other factors to initiate DNA repair. Serine is the major residue for ADP-ribosylation upon DNA damage, which strictly depends on HPF1. Here, we report the crystal structures of human HPF1/PARP1-CAT ΔHD complex at 1.98 Å resolution, and mouse and human HPF1 at 1.71 Å and 1.57 Å resolution, respectively. Our structures and mutagenesis data confirm that the structural insights obtained in a recent HPF1/PARP2 study by Suskiewicz et al. apply to PARP1. Moreover, we quantitatively characterize the key residues necessary for HPF1/PARP1 binding. Our data show that through salt-bridging to Glu284/Asp286, Arg239 positions Glu284 to catalyze serine ADP-ribosylation, maintains the local conformation of HPF1 to limit PARP1 automodification, and facilitates HPF1/PARP1 binding by neutralizing the negative charge of Glu284. These findings, along with the high-resolution structural data, may facilitate drug discovery targeting PARP1.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ADP-Ribosylation
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Amino Acid Sequence
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Animals
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Binding Sites
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Carrier Proteins / chemistry*
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Carrier Proteins / genetics
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Carrier Proteins / metabolism
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Cloning, Molecular
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Crystallography, X-Ray
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DNA / chemistry*
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DNA / genetics
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DNA / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Gene Expression
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Genetic Vectors / chemistry
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Genetic Vectors / metabolism
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Glutamine / metabolism
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Histones / chemistry*
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Histones / genetics
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Histones / metabolism
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Humans
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Mice
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Models, Molecular
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Nuclear Proteins / chemistry*
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism
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Poly (ADP-Ribose) Polymerase-1 / chemistry*
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Poly (ADP-Ribose) Polymerase-1 / genetics
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Poly (ADP-Ribose) Polymerase-1 / metabolism
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Protein Binding
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Protein Conformation, alpha-Helical
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Protein Conformation, beta-Strand
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Protein Interaction Domains and Motifs
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Protein Isoforms / chemistry
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Sequence Alignment
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Sequence Homology, Amino Acid
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Serine / metabolism*
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Static Electricity
Substances
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Carrier Proteins
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HPF1 protein, human
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Histones
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Nuclear Proteins
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Protein Isoforms
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Recombinant Proteins
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Glutamine
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Serine
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DNA
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PARP1 protein, human
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Poly (ADP-Ribose) Polymerase-1