DNA-dependent SUMO modification of PARP-1

DNA Repair (Amst). 2013 Sep;12(9):761-73. doi: 10.1016/j.dnarep.2013.07.001. Epub 2013 Jul 18.

Abstract

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.

Keywords: DNA binding; DNA repair; PARP-1; Posttranslational modification; SUMO; Transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Catalytic Domain
  • Chromatin / chemistry
  • Chromatin / physiology
  • DNA / chemistry
  • DNA / physiology*
  • DNA Breaks
  • DNA-Binding Proteins / chemistry
  • Enzyme Activation
  • HEK293 Cells
  • Humans
  • Inverted Repeat Sequences
  • Nucleic Acid Conformation
  • Plasmids / chemistry
  • Plasmids / physiology
  • Poly (ADP-Ribose) Polymerase-1
  • Poly(ADP-ribose) Polymerases / chemistry
  • Poly(ADP-ribose) Polymerases / metabolism*
  • Protein Binding
  • SUMO-1 Protein / metabolism*
  • Sumoylation*
  • Ubiquitin-Conjugating Enzymes / chemistry

Substances

  • Chromatin
  • DNA-Binding Proteins
  • SUMO-1 Protein
  • DNA
  • Ubiquitin-Conjugating Enzymes
  • PARP1 protein, human
  • Poly (ADP-Ribose) Polymerase-1
  • Poly(ADP-ribose) Polymerases
  • ubiquitin-conjugating enzyme UBC9