Identification and Characterization of a Mammalian 39-kDa Poly(ADP-ribose) Glycohydrolase

Oka, Shunya; Katō, Jirō; Moss, Joel

doi:10.1074/jbc.m510290200

Cited by 259 publications

(321 citation statements)

References 44 publications

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“…In the case of PARG, there are about 50 000 molecules in mammalian cells [14,33], and following the 90 % silencing achieved with the present protocol, about 5000 PARG molecules would escape down-regulation. In addition ADP-ribose-(arginine) protein hydrolase 3 (AHR3) catalyzes an alternative enzymatic activity for the degradation of poly(ADP-ribose) and must therefore be taken into account for residual PAR degradation activity within PARG-silenced cells [34]. However, at this level, we see a dramatic accumulation of PAR molecules (Fig.…”

Section: Discussionmentioning

confidence: 92%

The roles of poly(ADP-ribose)-metabolizing enzymes in alkylation-induced cell death

Cohausz

Blenn

Malanga

et al. 2008

Cell. Mol. Life Sci.

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Abstract. Poly(ADP-ribose) (PAR) has been identified as a DNA damage-inducible cell death signal upstream of apoptosis-inducing factor (AIF). PAR causes the translocation of AIF from mitochondria to the nucleus and triggers cell death. In living cells, PAR molecules are subject to dynamic changes pending on internal and external stress factors. Using RNA interference (RNAi), we determined the roles of poly(ADP-ribose) polymerases-1 and -2 (PARP-1, PARP-2) and poly(ADP-ribose) glycohydrolase (PARG), the key enzymes configuring PAR molecules, in cell death induced by an alkylating agent. We found that PARP-1, but not PARP-2 and PARG, contributed to alkylation-induced cell death. Likewise, AIF translocation was only affected by PARP-1. PARP-1 seems to play a major role configuring PAR as a death signal involving AIF translocation regardless of the death pathway involved.

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Section: Discussionmentioning

confidence: 92%

The roles of poly(ADP-ribose)-metabolizing enzymes in alkylation-induced cell death

Cohausz

Blenn

Malanga

et al. 2008

Cell. Mol. Life Sci.

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“…tankyrases, reviewed in 22) that take place outside the nucleus. Hydrolysis of PAR chains created by other PARPs is likely to involve PARG or ARH3 23 . It is possible that these PARG inhibitors prevent such processes but modification of this assay would have to be undertaken to detect non-nuclear PAR.…”

Section: Discussionmentioning

confidence: 99%

An assay to measure poly(ADP ribose) glycohydrolase (PARG) activity in cells

et al. 2016

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After a DNA damage signal multiple polymers of ADP ribose attached to poly(ADP) ribose (PAR) polymerases (PARPs) are broken down by the enzyme poly(ADP) ribose glycohydrolase (PARG). Inhibition of PARG leads to a failure of DNA repair and small molecule inhibition of PARG has been a goal for many years. To determine whether biochemical inhibitors of PARG are active in cells we have designed an immunofluorescence assay to detect nuclear PAR after DNA damage. This 384-well assay is suitable for medium throughput high-content screening and can detect cell-permeable inhibitors of PARG from nM to µM potency. In addition, the assay has been shown to work in murine cells and in a variety of human cancer cells. Furthermore, the assay is suitable for detecting the DNA damage response induced by treatment with temozolomide and methylmethane sulfonate (MMS). Lastly, the assay has been shown to be robust over a period of several years.

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“…We previously reported that ARH3 possesses poly(ADPr) glycohydrolase activity (10). The linkage hydrolyzed by ARH3 and ARH1 in poly(ADP-ribose) is also stereospecific at the C-1Љ position with the ␣-anomer present in the polymer (Fig.…”

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confidence: 92%

“…Three known members (ARH1-3) of the ARH family of proteins are similar in molecular size (ϳ39 kDa) and amino acid sequence (10). As noted above, ARH1 catalyzes the hydrolysis of ADP-ribose-arginine and also hydrolyzes ADP-ribose linkages to guanidine.…”

mentioning

confidence: 95%

Hydrolysis of O-Acetyl-ADP-ribose Isomers by ADP-ribosylhydrolase 3

Kasamatsu

Nakao

Smith

et al. 2011

Journal of Biological Chemistry

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O-Acetyl-ADP-ribose (OAADPr), produced by the Sir2-catalyzed NAD ؉ -dependent histone/protein deacetylase reaction, regulates diverse biological processes. Interconversion between two OAADPr isomers with acetyl attached to the C-2؆ and C-3؆ hydroxyl of ADP-ribose (ADPr) is rapid. We reported earlier that ADP-ribosylhydrolase 3 (ARH3), one of three ARH proteins sharing structural similarities, hydrolyzed OAADPr to ADPr and acetate, and poly(ADPr) to ADPr monomers. ARH1 also hydrolyzed OAADPr and poly(ADPr) as well as ADP-ribose-arginine, with arginine in ␣-anomeric linkage to C-1؆ of ADP-ribose. Because both ARH3-and ARH1-catalyzed reactions involve nucleophilic attacks at the C-1؆ position, it was perplexing that the ARH3 catalytic site would cleave OAADPr at either the 2؆-or 3؆-position, and we postulated the existence of a third isomer, 1؆-OAADPr, in equilibrium with 2؆-and 3؆-isomers. A third isomer, consistent with 1؆-OAADPr, was identified at pH 9.0. Further, ARH3 OAADPr hydrolase activity was greater at pH 9.0 than at neutral pH where 3؆-OAADPr predominated. Consistent with our hypothesis, IC 50 values for ARH3 inhibition by 2؆-and 3؆-N-acetyl-ADPr analogs of OAADPr were significantly higher than that for ADPr. ARH1 also hydrolyzed OAADPr more rapidly at alkaline pH, but cleavage of ADP-ribose-arginine was faster at neutral pH than pH 9.0. ARH3-catalyzed hydrolysis of OAADPr in H 218 O resulted in incorporation of one 18 O into ADP-ribose by mass spectrometric analysis, consistent with cleavage at the C-1؆ position. Together, these data suggest that ARH family members, ARH1 and ARH3, catalyze hydrolysis of the 1؆-O linkage in their structurally diverse substrates.Mono-ADP-ribosylation is a post-translational modification, in which the ADP-ribose (ADPr) 5 moiety of NAD is transferred to an acceptor protein (1). This modification serves as the mechanism by which several bacterial toxins (e.g. Pseudomonas exoenzyme S, cholera toxin, diphtheria toxin) exert their effects on mammalian cells (2, 3). Mammalian cells also produce endogenous ADP-ribosyltransferases that catalyze reactions similar to the bacterial toxins, specifically, the ADPribosylation of arginine residues in proteins (4). In addition, mammalian cells possess hydrolases that cleave the ADPr-protein linkage, releasing ADPr and regenerating the unmodified protein (5, 6). An ADP-ribosyl(arginine) hydrolase, termed ARH1, catalyzes in a stereospecific manner, hydrolysis of the ␣-linkage of arginine-ribose found in ADP-ribosyl(arginine)-protein to ADPr and (arginine)-protein (7, 8), consistent with the regulation of ADP-ribosyl(arginine)-protein levels by opposing activities of transferases and hydrolases, participating in an ADP-ribosylation cycle (4, 9).Three known members (ARH1-3) of the ARH family of proteins are similar in molecular size (ϳ39 kDa) and amino acid sequence (10). As noted above, ARH1 catalyzes the hydrolysis of ADP-ribose-arginine and also hydrolyzes ADP-ribose linkages to guanidine. The reaction is stereospecific, and only the ␣-ano...

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Identification and Characterization of a Mammalian 39-kDa Poly(ADP-ribose) Glycohydrolase

Abstract: , are located in a region similar to that required for activity in ARH1 but different from the location of the critical vicinal glutamates in the PARG catalytic site. All findings are consistent with the conclusion that ARH3 has PARG activity but is structurally unrelated to PARG.

Cited by 259 publications

References 44 publications

The roles of poly(ADP-ribose)-metabolizing enzymes in alkylation-induced cell death

The roles of poly(ADP-ribose)-metabolizing enzymes in alkylation-induced cell death

An assay to measure poly(ADP ribose) glycohydrolase (PARG) activity in cells

Hydrolysis of O-Acetyl-ADP-ribose Isomers by ADP-ribosylhydrolase 3

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