Characterization of polymers of adenosine diphosphate ribose generated in vitro and in vivo

Alvarez-Gonzalez, Rafael; Jacobson, Myron K.

doi:10.1021/bi00385a042

Cited by 229 publications

(184 citation statements)

References 30 publications

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“…How the polymer branching impacts on these parameters is not known at present. However, in general, branching increases with PAR size [36,37]. We identified PARP-1 as the major configurator of PAR as a cell death signal, while …”

Section: Discussionmentioning

confidence: 83%

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: 83%

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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“…An aqueous solution of PAR was HPLC-fractionated consulting a DNA Pac PA100 anion-exchange column (DIONEX) using a NaCl gradient. Modified sequencing gels for PAR quality check were carried out as described 69 using GELCODE Color silver stain (Pierce, Thermo-Fisher Scientific, Bonn, Germany).…”

Section: Methodsmentioning

confidence: 99%

Poly(ADP-ribose) binding to Chk1 at stalled replication forks is required for S-phase checkpoint activation

et al. 2013

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Damaged replication forks activate poly(ADP-ribose) polymerase 1 (PARP1), which catalyses poly(ADP-ribose) (PAR) formation; however, how PARP1 or poly(ADP-ribosyl)ation is involved in the S-phase checkpoint is unknown. Here we show that PAR, supplied by PARP1, interacts with Chk1 via a novel PAR-binding regulatory (PbR) motif in Chk1, independent of ATR and its activity. iPOND studies reveal that Chk1 associates readily with the unperturbed replication fork and that PAR is required for efficient retention of Chk1 and phosphorylated Chk1 at the fork. A PbR mutation, which disrupts PAR binding, but not the interaction with its partners Claspin or BRCA1, impairs Chk1 and the S-phase checkpoint activation, and mirrors Chk1 knockdown-induced hypersensitivity to fork poisoning. We find that long chains, but not short chains, of PAR stimulate Chk1 kinase activity. Collectively, we disclose a previously unrecognized mechanism of the S-phase checkpoint by PAR metabolism that modulates Chk1 activity at the replication fork.

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“…For example, poly(ADP) ribosylation (PARylation) plays a particularly important role in base excision repair with poly(ADP) ribose (PAR) polymerase 1 (PARP1) detecting single strand breaks that occur during this pathway. PARP1, which binds to these single strand breaks undergoes auto-modification creating up to 200 PAR chains 1 , that subsequently recruit the rest of the repair machinery including XRCC1 and POLB to complete the repair. The ADP ribose chains on PARP are hydrolysed by the enzyme poly(ADP) glycohydrolase (PARG).…”

Section: Introductionmentioning

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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Characterization of polymers of adenosine diphosphate ribose generated in vitro and in vivo

Cited by 229 publications

References 30 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

Poly(ADP-ribose) binding to Chk1 at stalled replication forks is required for S-phase checkpoint activation

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

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