Expression and activity of poly(ADP-ribose) glycohydrolase in cultured astrocytes, neurons, and C6 glioma cells

Sevigny, Mary B.; Silva, Jillian M.; Lan, Wen-Chun; Alano, Conrad C.; Swanson, Raymond A.

doi:10.1016/s0169-328x(03)00325-5

Cited by 10 publications

(20 citation statements)

References 38 publications

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“…We also demonstrate that PARP-1 and PARG reside in different subcellular compartments in non-neuronal tissue including the heart, testis, and kidney, where PARP-1 occurs primarily in the nucleus while PARG occurs mainly in the mitochondrial/cytosolic fraction. In agreement with our observations, Sevigny et al (Sevigny et al, 2003) demonstrated that in neuronal cultures, full length PARG was present only in the post-nuclear fraction where higher PARG activity was observed. Recent observations showed that full length PARG overexpressed in NIH 3T3 cells is localized exclusively in the nucleus during interphase and that it shuttles between nucleus and cytoplasm during the cell cycle (Ohashi et al, 2003).…”

Section: Discussionsupporting

confidence: 94%

“…We found that in rat brain, the full length PARG is the predominant form of the active enzyme ( Figure 1D). Interestingly, Sevigny et al, (2003) showed by Western blot analysis that primary neuronal cultures express mainly a short 63 kDa form of PARG. The differences observed could be associated with the different PARG antibodies used and possibly to their preferential affinity for PARG full length or the shorter forms of PARG.…”

Section: Discussionmentioning

confidence: 99%

“…However, very little is known about the anatomical or subcellular distribution of PARG in the brain, and nothing is yet known regarding its spatial and functional relationship to PARP-1. The message for PARG is present in the brain (Shimokawa et al, 1999) and active PARG is present in cultured neurons and astrocytes (Sevigny et al, 2003). The purification of PARG from nuclear and post-nuclear extracts suggests that PARG could be localized both in the cytoplasmic and the nuclear compartments.…”

Section: Introductionmentioning

confidence: 99%

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Spatial and functional relationship between poly(ADP-ribose) polymerase-1 and poly(ADP-ribose) glycohydrolase in the brain

Poitras

Koh

et al. 2007

Neuroscience

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Poly(ADP-ribose) polymerases (PARPs) are members of a family of enzymes that utilize NAD + as substrate to form large ADP-ribose polymers (PAR) in the nucleus. PAR has a very short half life due to its rapid degradation by poly(ADP-ribose) glycohydrolase (PARG). PARP-1 mediates acute neuronal cell death induced by a variety of insults including cerebral ischemia, MPTP-induced Parkinsonism, and CNS trauma. While PARP-1 is localized to the nucleus, PARG resides in both the nucleus and cytoplasm. Surprisingly, there appears to be only one gene encoding PARG activity, which has been characterized in vitro to generate different splice variants, in contrast to the growing family of PARPs. Little is known regarding the spatial and functional relationships of PARG and PARP-1. Here we evaluate PARG expression in the brain and its cellular and subcellular distribution in relation to PARP-1. Anti-PARG (α-PARG) antibodies raised in rabbits using a purified 30 kDa C-terminal fragment of murine PARG recognize a single band at 111 kDa in the brain. Western blot analysis also shows that PARG and PARP-1 are evenly distributed throughout the brain. Immunohistochemical studies using α-PARG antibodies reveal punctate cytosolic staining, whereas anti-PARP-1 (α-PARP-1) antibodies demonstrate nuclear staining. PARG is enriched in the mitochondrial fraction together with manganese superoxide dismutase (MnSOD) and cytochrome C (Cyt C) following whole brain subcellular fractionation and Western blot analysis. Confocal microscopy confirms the co-localization of PARG and Cyt C. Finally, PARG translocation to the nucleus is triggered by NMDA-induced PARP-1 activation. Therefore, the subcellular segregation of PARG in the mitochondria and PARP-1 in the nucleus suggests that PARG translocation is necessary for their functional interaction. This translocation is PARP-1 dependent, further demonstrating a functional interaction of PARP-1 and PARG in the brain.

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Section: Discussionsupporting

confidence: 94%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Spatial and functional relationship between poly(ADP-ribose) polymerase-1 and poly(ADP-ribose) glycohydrolase in the brain

Poitras

Koh

et al. 2007

Neuroscience

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“…Activity of recombinant PARP-1 was measured as described (46,47). In brief, the reaction mixture contained 50 mM Tris⅐HCl (pH 8.0), 10 mM MgCl 2 , 10 mM DTT, 10% glycerol, 25 g͞ml calf thymus DNA, 25 g͞ml histones, 210 M NAD ϩ , 25 pCi 14 C-NAD ϩ (248 mCi͞mmol, Amersham Pharmacia), 4 units of purified recombinant PARP-1 (Trevigen), and tetracycline derivatives or PARP inhibitors at the designated concentrations.…”

Section: Methodsmentioning

confidence: 99%

Minocycline inhibits poly(ADP-ribose) polymerase-1 at nanomolar concentrations

Alano

Kauppinen

Valls

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

223

181

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Poly(ADP-ribose) polymerase-1 (PARP-1), when activated by DNA damage, promotes both cell death and inflammation. Here we report that PARP-1 enzymatic activity is directly inhibited by minocycline and other tetracycline derivatives that have previously been shown to have neuroprotective and anti-inflammatory actions. These agents were evaluated by using cortical neuron cultures in which PARP-1 activation was induced by the genotoxic agents N-methyl-N -nitro-N-nitrosoguanidine (MNNG) or 3-morpholinosydnonimine (SIN-1). In both conditions, neuronal death was reduced by >80% either by 10 M 3,4-dihydro-5-[4-(1-piperidinyl)butoxy]-1(2H)-isoquinolinone, an established PARP inhibitor, or by 100 nM minocycline. Neuronal NAD ؉ depletion and poly(ADP-ribose) formation, which are biochemical markers of PARP-1 activation, were also blocked by 100 nM minocycline. A direct, competitive inhibition of PARP-1 by minocycline (K i ‫؍‬ 13.8 ؎ 1.5 nM) was confirmed by using recombinant PARP-1 in a cell-free assay. Comparison of several tetracycline derivatives showed a strong correlation (r 2 ‫؍‬ 0.87) between potency as a PARP-1 inhibitor and potency as a neuroprotective agent during MNNG incubations, with the rank order of potency being minocycline > doxycycline > demeclocycline > chlortetracycline. These compounds are known to have other actions that could contribute their neuroprotective effects, but at far higher concentrations than shown here to inhibit PARP-1. The neuroprotective and antiinflammatory effects of minocycline and other tetracycline derivatives may be attributable to PARP-1 inhibition in some settings.death ͉ doxycycline ͉ inflammation ͉ neuron M inocycline and other tetracycline derivatives have neuroprotective effects unrelated to their antimicrobial properties (1). Minocycline can reduce neuronal death after excitotoxicity and ionizing radiation in culture (2, 3), and in animal models of stroke (1, 3-5), Parkinson's disease (6, 7), Huntington's disease (8), and amyotrophic lateral sclerosis (9). The neuroprotective effects of minocycline have been attributed both to reduced inflammation and a direct effect on neuronal survival (2, 3, 5, 9, 10).Poly(ADP-ribose) polymerase-1 (PARP-1) plays a key role in neuronal death and survival under stress conditions (11). PARP-1 is the most abundant of several PARP family members, accounting for Ͼ85% of nuclear PARP activity (11). When activated by DNA damage, PARP-1 consumes NAD ϩ to form branched poly(ADP-ribose) on target proteins. Poly(ADPribose) formation on histones and enzymes involved in DNA repair appears to facilitate DNA repair by preventing chromatid exchange and by loosening histone wrapping (12, 13). Poly(ADPribose) formation also has effects on gene transcription through interactions with transcription factors, notably NF-B (14-16), and PARP-1 inhibition or gene deletion attenuates the brain microglial response to cytokines and other triggers (17-19). Extensive PARP-1 activation can, in addition, lead to neuronal death through mechanisms linked to NAD ϩ deplet...

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“…An arrest of the embryological development phase in PARG-/-embryos was due to an increase in PAR which prevented cellular replication characterized by unhatched blastocysts [6] and in C6 glioma tumor dividing cells there was a high PARG activity therefore, lack of PARG could anticipate cell growth arrest [45], showing that PARG protein deficiency may have a crucial role in tumor treatment.…”

Section: Action Of Parp and Parg In Cell Proliferation And Differentimentioning

confidence: 99%

PARP and PARG Inhibitors—New Therapeutic Targets in Cancer Treatment

Fauzee

Perucho

Wang

2010

Pathol. Oncol. Res.

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Today, the number of cancer patients throughout the world is increasing alarmingly and as per the World Health Organisation (WHO) data and statistics the prediction for the year 2020 will be 15 million new cases as compared to only 10 million cases in year 2000 leaving us dumbfounded. A lot of effort has been put in by researchers and scientists over decades to find drugs helpful in the treatment of cancers for the benefit of patients--the latest being the Poly ADP-ribose polymerase (PARP) and the Poly ADP-ribose glycohydrolase (PARG) inhibitors. This review highlights their mechanism of action under the rationale of their use and current development in the field of cancer.

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Expression and activity of poly(ADP-ribose) glycohydrolase in cultured astrocytes, neurons, and C6 glioma cells

Cited by 10 publications

References 38 publications

Spatial and functional relationship between poly(ADP-ribose) polymerase-1 and poly(ADP-ribose) glycohydrolase in the brain

Spatial and functional relationship between poly(ADP-ribose) polymerase-1 and poly(ADP-ribose) glycohydrolase in the brain

Minocycline inhibits poly(ADP-ribose) polymerase-1 at nanomolar concentrations

PARP and PARG Inhibitors—New Therapeutic Targets in Cancer Treatment

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