Proteolysis of poly(ADP-ribose) polymerase by caspase 3: Kinetics of cleavage of mono(ADP-ribosyl)ated and DNA-bound substrates

Damien D'Amours, Marc Germain, Kim Orth, Vishva M. Dixit, Guy G. Poirier

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Poly(ADP-ribose) polymerase (PARP) is an abundant nuclear enzyme which is responsible for synthesis of poly(ADP-ribose) in response to DNA damage caused by numerous agents and during DNA base excision repair. After DNA damage, the enzyme binds to nicks in DNA through its N-terminal zinc fingers and catalyzes the formation of poly(ADP-ribose) on various nuclear acceptors including itself. When DNA damage is extensive, cells induce their own demise by activating the proteases that induce apoptosis (caspases) which cleave PARP and other death substrates. Here we report the development of a new approach to investigate the sensitivity of mono(ADP-ribosyl)ated and DNA- bound PARP to cleavage during apoptosis. The development of a stoichiometric labeling procedure of the enzyme has allowed us to evaluate the catalytic properties of caspase 3 toward mono(ADP-ribosyl)ated PARP at various enzyme:substrate molar ratios. We show that low levels of automodification (≤3 U of ADP-ribose per chain) do not inhibit the proteolysis of the substrate. In addition, we demonstrate that binding of unmodified PARP to DNA influences the kinetics of its cleavage by caspase 3.

Original languageEnglish (US)
Pages (from-to)3-10
Number of pages8
JournalRadiation Research
Volume150
Issue number1
DOIs
StatePublished - Jul 1998

Fingerprint

ribose
NAD ADP-ribosyltransferase
adenosine diphosphate
Poly(ADP-ribose) Polymerases
caspase-3
Caspase 3
proteolysis
Adenosine Diphosphate
Proteolysis
cleavage
deoxyribonucleic acid
kinetics
DNA damage
DNA
DNA Damage
Poly Adenosine Diphosphate Ribose
Enzymes
enzymes
apoptosis
Adenosine Diphosphate Ribose

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Radiology Nuclear Medicine and imaging
  • Biophysics
  • Radiation

Cite this

Proteolysis of poly(ADP-ribose) polymerase by caspase 3 : Kinetics of cleavage of mono(ADP-ribosyl)ated and DNA-bound substrates. / D'Amours, Damien; Germain, Marc; Orth, Kim; Dixit, Vishva M.; Poirier, Guy G.

In: Radiation Research, Vol. 150, No. 1, 07.1998, p. 3-10.

Research output: Contribution to journalArticle

D'Amours, Damien ; Germain, Marc ; Orth, Kim ; Dixit, Vishva M. ; Poirier, Guy G. / Proteolysis of poly(ADP-ribose) polymerase by caspase 3 : Kinetics of cleavage of mono(ADP-ribosyl)ated and DNA-bound substrates. In: Radiation Research. 1998 ; Vol. 150, No. 1. pp. 3-10.
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