Deletion Mutants of Poly(ADP-Ribose) Polymerase A Support a Model of Cyclic Association and Dissociation of Enzyme from DNA Ends During DNA Repair

Smulson, Mark E.; Istock, Nancy L.; Ding, Ruchuang; Cherney, Barry

doi:10.1021/bi00186a018

Cited by 57 publications

(38 citation statements)

References 26 publications

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“…The discrepancies in the possible role of ADPRT in DNA repair may be attributable to the fact that previous studies used either ADPRT inhibitors, which may have additional nonspecific effects, or spontanous ADPRT mutant cell lines, which contain reduced levels of ADPRT. In agreement with our data showing that ADPRT is not a critical regulatory component in the DNA repair process are studies demonstrating that DNA repair was apparently independent of ADPRT (Satoh and Lindahl 1992;Satoh et al 1993;Smulson et al 1994). These results suggest that ADPRT acts as a "nick-protection" molecule to prevent accidental homologous recombination events and is needed for the maintenance of chromatin structure (Althaus et al 1990;Satoh et al 1994).…”

Section: Dna Repair Processes Are Not Affected By the Lack Of Poly(adsupporting

confidence: 92%

Mice lacking ADPRT and poly(ADP-ribosyl)ation develop normally but are susceptible to skin disease.

Wang¹,

Auer²,

Stingl³

et al. 1995

Genes Dev.

743

504

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Poly(ADP-ribosyl)ation is catalyzed by NAD + :protein(ADP-ribosyl)transferase (ADPRT), a chromatinassociated enzyme which, in the presence of DNA breaks, transfers ADP-ribose from NAD + to nuclear proteins. This post-translational modification has been implicated in many fundamental processes, like DNA repair, chromatin stability, cell proliferation, and cell death. To elucidate the biological function of ADPRT and poly(ADP-ribosyl)ation in vivo the gene was inactivated in the mouse germ line. Mice homozygous for the ADPRT mutation are healthy and fertile. Analysis of mutant tissues and fibroblasts isolated from mutant fetuses revealed the absence of ADPRT enzymatic activity and poly(ADP-ribose), implying that no poly(ADP-ribosyl)ated proteins are present. Mutant embryonic fibroblasts were able to efficiently repair DNA damaged by UV and alkylating agents. However, proliferation of mutant primary fibroblasts as well as thymocytes following y-radiation in vivo was impaired. Moreover, mutant mice are susceptible to the spontaneous development of skin disease as -30% of older mice develop epidermal hyperplasia. The generation of viable ADPRT-/-mice negates an essential role for this enzyme in normal chromatin function, but the impaired proliferation and the onset of skin lesions in older mice suggest a function for ADPRT in response to environmental stress.

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Section: Dna Repair Processes Are Not Affected By the Lack Of Poly(adsupporting

confidence: 92%

Mice lacking ADPRT and poly(ADP-ribosyl)ation develop normally but are susceptible to skin disease.

Wang¹,

Auer²,

Stingl³

et al. 1995

Genes Dev.

743

504

View full text Add to dashboard Cite

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“…Cell lines transfected with and overproducing the 'DNA binding domain' (DBD) of PADPRP, thus inhibiting endogenous PADPRP activation, are also hypersensitive to monofunctional alkylating agents, and are unable to carry out unscheduled DNA synthesis (Molinette et al, 1993). Similar results have been obtained by reducing endogenous PADPRP synthesis by the use of antisense olignucleotides to PADPRP (Smulson et al, 1994). Finally, elegant in vitro experiments using crude cell extracts which can carry out DNA repair have established that PADPRP, in the absence of substrate, blocks DNA strand breaks snd prevents subsequent steps leading to religation of the DNA (Satoh and Lindhal, 1992;Satoh et al, 1993).…”

mentioning

confidence: 72%

Potentiation of temozolomide-induced cytotoxicity: a comparative study of the biological effects of poly(ADP-ribose) polymerase inhibitors

Boulton¹,

Pemberton²,

Porteous³

et al. 1995

Br J Cancer

121

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Summary Four poly(ADP-ribose) polymerase (PADPRP) inhibitors [3-aminobenzamide, benzamide, 3,4-dihydro-5-methoxyisoquinolin-1(2H)-one (PD 128763) and 8-hydroxy-2-methylquinazolin-4(3H)-one (NU1025)] were compared with respect to their effects on a number of biological end points. The following parameters were assessed: their ability to inhibit the enzyme in permeabilised L1210 cells; their ability to potentiate the cytotoxicity of temozolomide (including the cytotoxicity of the compounds per se); their ability to increase net levels of temozolomide-induced DNA strand breaks and inhibit temozolomide-induced NAD depletion. PD 128763 and NU1025 were equipotent as PADPRP inhibitors, and 40-and 50-fold more potent than benzamide and 3-aminobenzamide respectively. All the compounds acted in a concentration-dependent manner to potentiate the cytotoxicity and increase DNA strand break levels in cells treated with temozolomide. There was an excellent correlation between the potency of the compounds as PADPRP inhibitors and their effects on cell survival and DNA repair. Temozolomide treatment caused a decrease in cellular NAD levels, and this was abolished by the PADPRP inhibitors. In conclusion, the new generation of PADPRP inhibitors are at least 50-fold more effective than 3-aminobenzamide as chemopotentiators, and can be used at micromolar rather than millimolar concentrations in intact cells.

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“…Due to the charge repulsion the protein rapidly dissociates from DNA (4,33,34). Therefore, we next tested the ability of PARP to bind hairpin-containing DNA under conditions conducive to PARP automodification.…”

Section: Results and Disscussionmentioning

confidence: 99%

Transcriptional Repression by Binding of Poly(ADP-ribose) Polymerase to Promoter Sequences

Soldatenkov¹,

Chasovskikh²,

Potaman³

et al. 2002

Journal of Biological Chemistry

104

106

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Poly(ADP-ribose) polymerase (PARP) is a DNA-binding enzyme that plays roles in response to DNA damage, apoptosis, and genetic stability. Recent evidence has implicated PARP in transcription of eukaryotic genes. However, the existing paradigm tying PARP function to the presence of DNA strand breaks does not provide a mechanism by which it may be recruited to gene-regulating domains in the absence of DNA damage. Here we report that PARP can bind to the DNA secondary structures (hairpins) in heteroduplex DNA in a DNA end-independent fashion and that automodification of PARP in the presence of NAD ؉ inhibited its hairpin binding activity. Atomic force microscopic images show that in vitro PARP protein has a preference for the promoter region of the PARP gene in superhelical DNA where the dyad symmetry elements likely form hairpins according to DNase probing. Using a chromatin cross-linking and immunoprecipitation assay we show that PARP protein binds to the chromosomal PARP promoter in vivo. Reporter gene assays have revealed that the transcriptional activity of the PARP promoter is 4 -5-fold greater in PARP knockout cells than in wild type fibroblasts. Reintroduction of vectors expressing full-length PARP protein or its truncated mutant (DNA-binding domain retained but lacking catalytic activity) into PARP ؊/؊ cells has conferred transcriptional down-regulation of the PARP gene promoter. These data provide support for PARP protein as a potent regulator of transcription including down-regulation of its own promoter.Poly(ADP-ribose) polymerase (PARP, 1 EC 2.4.2.30) is a chromatin-associated enzyme that catalyzes the transfer of successive units of the ADP-ribose moiety from NAD ϩ to itself and other nuclear acceptor proteins (1). PARP is a zinc fingercontaining protein, which allows enzyme binding to either double or single strand DNA breaks without any apparent sequence preference (2, 3). The catalytic activity of PARP is strictly dependent on the presence of strand breaks in DNA and is modulated by the level of automodification (4,5). Data from many studies show that PARP is involved in numerous biological functions, all of which are associated with breaking and rejoining DNA strands, and it plays a pivotal role in DNA damage repair (2, 6 -8).Recent studies have implicated PARP in transcription of eukaryotic genes (9 -16). PARP-dependent gene regulation involves poly(ADP-ribosyl)ation of transcription factors, which, in turn, prevents their binding to specific promoter sequences (10). The basal transcription factors TFIIF and TEF-1 as well as transcription factors TATA box-binding protein, YY1, SP-1, cAMP-response element-binding protein, p53, and NFB are all highly specific substrates for poly(ADP-ribosyl)ation (10,11,14,16). PARP may also interact directly with gene promoters. For instance, recombinant full-length PARP bound the DNA sequences within the MCAT1 regulatory element (11) and to the DF4 protein binding site of the Pax-6 gene neuroretinaspecific enhancer (17). Furthermore, PARP involvement in the active...

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Deletion Mutants of Poly(ADP-Ribose) Polymerase A Support a Model of Cyclic Association and Dissociation of Enzyme from DNA Ends During DNA Repair

Cited by 57 publications

References 26 publications

Mice lacking ADPRT and poly(ADP-ribosyl)ation develop normally but are susceptible to skin disease.

Mice lacking ADPRT and poly(ADP-ribosyl)ation develop normally but are susceptible to skin disease.

Potentiation of temozolomide-induced cytotoxicity: a comparative study of the biological effects of poly(ADP-ribose) polymerase inhibitors

Transcriptional Repression by Binding of Poly(ADP-ribose) Polymerase to Promoter Sequences

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