Structure/function analysis of PARP-1 in oxidative and nitrosative stress-induced monomeric ADPR formation.
Poly adenosine diphosphate-ribose polymerase-1 (PARP-1) is a multifunctional enzyme that is involved in two major cellular responses to oxidative and nitrosative (O/N) stress: detection and response to DNA damage via formation of protein-bound poly adenosine diphosphate-ribose (PAR), and formation of the soluble 2(nd) messenger monomeric adenosine diphosphate-ribose (mADPR). Previous studies have delineated specific roles for several of PARP-1's structural domains in the context of its involvement in a DNA damage response. However, little is known about the relationship between the mechanisms through which PARP-1 participates in DNA damage detection/response and those involved in the generation of monomeric ADPR. To better understand the relationship between these events, we undertook a structure/function analysis of PARP-1 via reconstitution of PARP-1 deficient DT40 cells with PARP-1 variants deficient in catalysis, DNA binding, auto-PARylation, and PARP-1's BRCT protein interaction domain. Analysis of responses of the respective reconstituted cells to a model O/N stressor indicated that PARP-1 catalytic activity, DNA binding, and auto-PARylation are required for PARP-dependent mADPR formation, but that BRCT-mediated interactions are dispensable. As the BRCT domain is required for PARP-dependent recruitment of XRCC1 to sites of DNA damage, these results suggest that DNA repair and monomeric ADPR 2(nd) messenger generation are parallel mechanisms through which PARP-1 modulates cellular responses to O/N stress.
Buelow
Ben
B
Department of Pediatrics, University of Washington, Seattle, Washington, United States of America.
Uzunparmak
Burak
B
Paddock
Marcia
M
Scharenberg
Andrew M
AM
D000246:Adenosine Diphosphate Ribose / Q000096:biosynthesis*; D000818:Animals; D002384:Catalysis; D002460:Cell Line; D002645:Chickens; D004249:DNA Damage; D015538:Nitrosation; D018384:Oxidative Stress; D011065:Poly(ADP-ribose) Polymerases / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship
D000246:Adenosine Diphosphate Ribose; D011065:Poly(ADP-ribose) Polymerases