Modulation of DNA fragmentation factor 40 nuclease activity by poly(ADP-ribose) polymerase-1.
Poly(ADP-ribose) polymerase-1 (PARP-1) influences numerous cellular processes, including DNA repair, transcriptional regulation, and caspase-independent cell death, by utilizing NAD(+) to synthesize long chains of poly(ADP-ribose) (PAR) on target proteins, including itself. During the apoptotic response, caspases-3 and -7 cleave PARP-1, thereby inhibiting its activity. Here, we have examined the role of PARP-1 activation and cleavage in the latter stages of apoptosis in response to DNA fragmentation. PARP-1 poly(ADP-ribosyl)ation correlated directly with induction of apoptosis by the lipid peroxidation product, 4-hydroxy-2-nonenal. A significant decrease in PAR accumulation was observed upon caspase or DNA fragmentation factor 40 (DFF40) inhibition. Because DNA fragmentation mediated by DFF40 augmented PARP-1 modification status in apoptotic cells, we hypothesized that PARP-1 alters DFF40 function following PAR accumulation. Indeed, PARP-1, in the presence of NAD(+), significantly decreased DFF40 activity on plasmid substrates. Conversely, PARP-1 enhanced the DNase activity of DFF40 in the absence of NAD(+). The inhibition of DFF40 activity in the presence of NAD(+) was reduced by co-incubation with poly(ADP-ribose) glycohydrolase and a PARP inhibitor. Additionally, caspase-cleaved PARP-1, in the presence of NAD(+), did not inhibit DFF40 activity significantly. Our results suggest that PARP-1 poly(ADP-ribosyl)ation is a terminal event in the apoptotic response that occurs in response to DNA fragmentation and directly influences DFF40 activity.
West
James D
JD
Department of Biochemistry, Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology and the Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-0146, USA.
Ji
Chuan
C
Marnett
Lawrence J
LJ
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D053148:Caspase 3; D053179:Caspase 7; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D053938:DNA Fragmentation; D004260:DNA Repair; D003851:Deoxyribonucleases / Q000737:chemistry* / Q000378:metabolism; D004789:Enzyme Activation; D000939:Epitopes / Q000737:chemistry; D005822:Genetic Vectors; D006026:Glycoside Hydrolases / Q000737:chemistry; D006801:Humans; D047468:Immunoprecipitation; D015227:Lipid Peroxidation; D051379:Mice; D008954:Models, Biological; D008958:Models, Molecular; D010957:Plasmids / Q000378:metabolism; D011064:Poly Adenosine Diphosphate Ribose / Q000737:chemistry; D011065:Poly(ADP-ribose) Polymerases / Q000737:chemistry*; D000075223:Poly-ADP-Ribose Binding Proteins; D011485:Protein Binding; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection
D000939:Epitopes; D000075223:Poly-ADP-Ribose Binding Proteins; D011064:Poly Adenosine Diphosphate Ribose; D011065:Poly(ADP-ribose) Polymerases; C113696:DFFB protein, human; D003851:Deoxyribonucleases; D006026:Glycoside Hydrolases; C022407:poly ADP-ribose glycohydrolase; C505407:CASP3 protein, human; C505520:CASP7 protein, human; C505409:Casp3 protein, mouse; C505524:Casp7 protein, mouse; D053148:Caspase 3; D053179:Caspase 7; D020169:Caspases