Poly(ADP‐Ribose) Polymerase in Plant Nuclei

Abstract

We show that poly(ADP-ribose) polymerase is present in maize, pea and wheat nuclei. We have identified the enzyme product as poly(ADP-ribose) by purification and electrophoresis on a DNA sequencing gel. This reveals a polymer ladder consisting of up to 45 residues. The polymer product from maize, after digestion with snake venom phosphodiesterase, gave only 5'-AMP and (phosphoribosy1)-AMP; the mean chain length of the polymer was 5 and 11 residues in two separate experiments. The optimum pH of the plant enzyme is greater than pH 7.0 in pea, wheat and maize; the optimum temperature for enzyme activity is approximately 15 "C. The K, for NAD' for the enzyme from maize is estimated to be approximately 50 pM under optimal conditions. Several compounds (nicotinamide, deoxythymidine, 3-aminobenzamide, 3-methoxybenzamide and 5-bromodeoxyuridine) that specifically inhibit the animal enzyme also inhibit the enzyme from plants. The ratio of the IC,, for 5-bromodeoxyuridine to the IC,, for 3-aminobenzamide in maize is similar to that of the animal enzyme indicating that the enzyme involved is poly(ADP-ribose) polymerase and not mono(ADP-ribosyl) transferase. SDS gel electrophoresis and gel filtration analysis of a crude extract of maize nuclei indicate a molecular mass for poly(ADP-ribose) polymerase of approximately 114 kDa.

Disruption of Poly(ADP-ribosyl)ation Mechanisms Alters Responses of Arabidopsis to Biotic Stress

“…Since PARP consumes NAD + to synthesize ADP-Rib units, an examination of cellular NAD + levels can be used as an indirect measure of poly (ADP-Rib) synthesis activities (Chen et al, 1994;Du et al, 2003;De Block et al, 2005;Ishikawa et al, 2009). Whereas no significant change in NAD + levels was seen for seedlings treated with avirulent pathogen, flg22 and/or 3AB (data not shown), a statistically significant 40% to 50% decrease in NAD + compared with mock-infiltrated samples was observed in leaves 12 h after infiltration with virulent Pst DC3000 (Supplemental Fig.…”

“…Use of pharmacological PARP inhibitors is a common way of overcoming such potential functional redundancy and also allows conditional inactivation of PARP activity. 3-Aminobenzamide (3AB) is a widely used PARP inhibitor in both animal (Bryant et al, 2005;Beauchamp et al, 2009;Ding et al, 2009;Hernandez et al, 2009) and plant (Phillips and Hawkins, 1985;Berglund et al, 1996;Amor et al, 1998;Tian et al, 2000;Adams-Phillips et al, 2008;Ishikawa et al, 2009) studies and has been shown to inhibit plant PARP enzymatic activity (Chen et al, 1994;Babiychuk et al, 1998). 3AB has also been used to demonstrate the linkage between PARP and PARG activity in plants.…”

“…There is evidence that plant PARPs are structurally and functionally homologous to mammalian PARP proteins (Chen et al, 1994;O'Farrell, 1995;Babiychuk et al, 1998;Doucet-Chabeaud et al, 2001). DNA damage induced by ionizing radiation activates Arabidopsis PARP1 and PARP2 gene expression (Doucet-Chabeaud et al, 2001).…”

UV‐B‐ and oxidative stress‐induced increase in nicotinamide and trigonelline and inhibition of defensive metabolism induction by poly(ADP‐ribose)polymerase inhibitor in plant tissue

“…An important source of NIC release in response to stress is the DNA strand break-activated eukaryotic enzyme poly(-ADP-ribose)polymerase (PADPRP) [13,14]. PADPRP is present in nuclei of various plant species [15] and shows similarities to animal PADPRP [16]. In plants, NIC can be metabolized to N-glucose nicotinic acid or TRIG via nicotinic acid [17].…”

Centromere Proteins Cenpa, Cenpb, and Bub3 Interact with Poly(ADP-ribose) Polymerase-1 Protein and Are Poly(ADP-ribosyl)ated

“…Poly(ADP-ribose) polymerase-1 (PARP-1) 1 is a multifunctional enzyme that catalyzes the formation of poly(ADP-ribose) polymers on acceptor proteins involved in the maintenance of chromatin structure and DNA repair (31)(32)(33)(34)(35)(36)(37). The addition of poly(ADP-ribose) units makes the acceptor proteins more negatively charged, thus altering their structure, function, and binding properties to DNA (38 -40).…”