DNA-dependent SUMO modification of PARP-1
| Autor: | Stephen C. West, Chris T. Williamson, Helle D. Ulrich, Sebastian Eustermann, Rajvee Shah, David Neuhaus, Nicola Zilio |
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| Rok vydání: | 2013 |
| Předmět: |
HMG-box
DNA repair DNA damage Poly ADP ribose polymerase SUMO-1 Protein Poly (ADP-Ribose) Polymerase-1 SUMO protein PARP-1 SUMO enzymes Biochemistry Article Catalytic Domain Humans DNA binding Molecular Biology Polymerase Base Sequence biology DNA Breaks Inverted Repeat Sequences Sumoylation DNA Cell Biology Chromatin DNA-Binding Proteins Enzyme Activation HEK293 Cells SUMO Ubiquitin-Conjugating Enzymes biology.protein Nucleic Acid Conformation Posttranslational modification Poly(ADP-ribose) Polymerases Transcription Plasmids Protein Binding |
| Zdroj: | DNA Repair |
| ISSN: | 1568-7864 |
| DOI: | 10.1016/j.dnarep.2013.07.001 |
| Popis: | Highlights • We examine the sumoylation of PARP-1 in response to different DNA ligands. • We characterise the dynamics of the PARP-1 SUMO conjugate by NMR. • Sumoylation of PARP-1 is stimulated by binding to intact, but not to damaged DNA. • Sumoylation does not interfere with PARP-1's DNA binding or catalytic activity. • PARP-1 sumoylation and catalytic activation follow independent regulatory mechanisms. Poly(ADP-ribose) polymerase 1 (PARP-1) plays an important role in DNA repair, but also contributes to other aspects of nucleic acid metabolism, such as transcriptional regulation. Modification of PARP-1 with the small ubiquitin-related modifier (SUMO) affects its function as a transcriptional co-activator of hypoxia-responsive genes and promotes induction of the heat shock-induced HSP70.1 promoter. We now report that PARP-1 sumoylation is strongly influenced by DNA. Consistent with a function in transcription, we show that sumoylation in vitro is enhanced by binding to intact, but not to damaged DNA, in a manner clearly distinct from the mechanism by which DNA damage stimulates PARP-1's catalytic activity. An enhanced affinity of PARP-1 for the SUMO-conjugating enzyme Ubc9 upon binding to DNA is likely responsible for this effect. Sumoylation does not interfere with the catalytic or DNA-binding properties of PARP-1, and structural analysis reveals no significant impact of SUMO on the conformation of PARP-1's DNA-binding domain. In vivo, sumoylated PARP-1 is associated with chromatin, but the modification is not responsive to DNA damage and is not affected by PARP-1 catalytic activity. Our results suggest that PARP-1's alternative modes of DNA recognition serve as a means to differentiate between distinct aspects of the enzyme's function. |
| Databáze: | OpenAIRE |
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