Enzymological properties of poly(ADP-ribose)polymerase: characterization of automodification sites and NADase activity

Desmarais, Yvan; Ménard, Luc; Lagueux, Jean; Poirier, Guy G.

doi:10.1016/0167-4838(91)99007-f

Cited by 92 publications

(42 citation statements)

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“…Reprobing of the immunoblot with antibodies to E2F-1 revealed a B60-kDa band, distinct from the 4114 kDa band detected with anti-PAR (Figure 2 right panel), thus, indicating that the modified protein was not E2F-1. PARP-1 is the main acceptor in these reactions, attributable to the presence of up to 28 automodification sites in the protein (Kawaichi et al, 1981;Cherney et al, 1987;Desmarais et al, 1991). In contrast to E2F-1, we have demonstrated that GST-p53 is an effective substrate for PARP-1 under the same conditions (Simbulan- Rosenthal et al, 2001).…”

Section: Resultsmentioning

confidence: 65%

PARP-1 binds E2F-1 independently of its DNA binding and catalytic domains, and acts as a novel coactivator of E2F-1-mediated transcription during re-entry of quiescent cells into S phase

et al. 2003

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The transcription factor E2F-1 is implicated in the activation of S-phase genes as well as induction of apoptosis, and is regulated by interactions with Rb and by cell cycle-dependent alterations in E2F-1 abundance. We earlier demonstrated a pivotal role for poly(ADPribose) polymerase-1 (PARP-1) in the regulation of E2F-1 expression and promoter activity during S-phase re-entry when quiescent cells re-enter the cell cycle. We now investigate the putative mechanism(s) by which PARP-1 may upregulate E2F-1 promoter activity during S-phase re-entry. DNase-1 footprint assays with purified PARP-1 showed that PARP-1 did not directly bind the E2F-1 promoter in a sequence-specific manner. In contrast to p53, a positive acceptor in poly(ADP-ribosyl)ation reactions, E2F-1 was not poly(ADP-ribosyl)ated by wildtype PARP-1 in vitro, indicating that PARP-1 does not exert a dual effect on E2F-1 transcriptional activation. Protein-binding reactions and coimmunoprecipitation experiments with purified PARP-1 and E2F-1, however, revealed that PARP-1 binds to E2F-1 in vitro. More significantly, physical association of PARP-1 and E2F-1 in vivo also occurred in wild-type fibroblasts 5 h after re-entry into S phase, coincident with the increase in E2F-1 promoter activity and expression of E2F-1-responsive Sphase genes cyclin A and c-Myc. Mapping of the interaction domains revealed that full-length PARP-1 as well as PARP-1 mutants lacking either the catalytic active site or the DNA-binding domain equally bind E2F-1, whereas a PARP-1 mutant lacking the automodification domain does not, suggesting that the protein interaction site is located in this central domain. Finally, gel shift analysis with end-blocked E2F-1 promoter sequence probes verified that the binding of PARP-1 to E2F-1 enhances binding to the E2F-1 promoter, indicating that PARP-1 acts as a positive cofactor of E2F-1-mediated transcription.

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Section: Resultsmentioning

confidence: 65%

PARP-1 binds E2F-1 independently of its DNA binding and catalytic domains, and acts as a novel coactivator of E2F-1-mediated transcription during re-entry of quiescent cells into S phase

et al. 2003

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“…There are several glutamic acid residues in this domain that are suggested to be the site for covalent binding with poly(ADP-ribose) on PARP-1 activation. 19,20 However, other groups argue that individual lysine residues, not glutamic acid, serve as acceptor sites for ADP ribosylation in the AMD. 21 The PAR modification of glutamic residues in the NAD ϩ -binding domain and the DBD has also been noted.…”

Section: Parp-1 Structure Properties and Activationmentioning

confidence: 99%

“…21 The PAR modification of glutamic residues in the NAD ϩ -binding domain and the DBD has also been noted. 19,20 Irrespective of the site, automodification is accepted as a mechanism for regulating PARP-1 activity and control PAR synthesis 19 ; however, the exact impact and mechanisms of automodification on enzyme function need further investigation.…”

Section: Parp-1 Structure Properties and Activationmentioning

confidence: 99%

Signaling Mechanism of Poly(ADP-Ribose) Polymerase-1 (PARP-1) in Inflammatory Diseases

Garg

2011

The American Journal of Pathology

163

140

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Poly(ADP-ribosyl)ation, attaching the ADP-ribose polymer chain to the receptor protein, is a unique posttranslational modification. Poly(ADP-ribose) polymerase-1 (PARP-1) is a well-characterized member of the PARP family. In this review, we provide a general update on molecular structure and structure-based activity of this enzyme. However, we mainly focus on the roles of PARP-1 in inflammatory diseases. Specifically, we discuss the signaling pathway context that PARP-1 is involved in to regulate the pathogenesis of inflammation. PARP-1 facilitates diverse inflammatory responses by promoting inflammation-relevant gene expression, such as cytokines, oxidation-reductionrelated enzymes, and adhesion molecules. Excessive activation of PARP-1 induces mitochondria-associated cell death in injured tissues and constitutes another mechanism for exacerbating inflammation. There are many posttranslational protein modifications (eg, phosphorylation, acetylation, methylation, and ubiquitylation) that are involved in a wide scope of cellular processes, including chromatin remodeling, transcriptional regulation, and signal transmission response to extracellular stimulation. Poly(ADP-ribosyl)ation (PARylation) is one of such essential protein modifications, whereby polymers of ADP-ribose (PARs) are formed from donor NAD ϩ molecules and covalently attached via an ester linkage to glutamic acid and less commonly to aspartic acid or lysine of target proteins.1-3 The target proteins generally contain a PAR-binding consensus motif that frequently overlaps with a functional domain, such as a protein-or DNA-binding domain (DBD), and thus accounts for PAR modification, altering the functional properties of the targets. 4 The process of PARylation is catalyzed by the poly-(ADP-ribose) polymerase (PARP) family of enzymes that consists of 18 members. 5 The PARPs, historically known as poly(ADP-ribose) synthases and poly(ADP-ribose) transferases, 6,7 show different structure, cellular location, and functions. 8,9 Only two members of this family (ie, PARP-1 and PARP-2) are DNA damage related; and PARP-1, the best-understood member, is an abundant nuclear enzyme and accounts for at least 85% of the cellular PARP activity. 10Since the discovery of PAR synthesis and PARP-1 decades ago, 11,12 new discoveries have been consistently published related to their structure, property, and functions. PARP-1 is a multifunctional enzyme and has a key role in the spatial and temporal organization of DNA repair, thus maintaining genome integrity and facilitating cell survival. PARP-1 catalyzes the synthesis and attachment of highly negatively charged PARs to target proteins, including histones, topoisomerases, DNA helicases, and single-strand break repair and base excision repair factors; and facilitates relaxation of the chromatin superstructure, protein-protein interaction, and DNAbinding ability of the members of the DNA repair machinery. A recently published article 13 reviews the role of PARP-1 in DNA repair. In addition, the importance of poly-(ADP-...

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“…Glutamic acid and aspartic acid residues are most likely to be modified by PARlation, with known examples of histone H2B PARlated in vitro at glutamic acid residue 2, 20 histone H1 PARlated at glutamic acid residues 2, 14, and the COOHterminal lysine, 21 and PARP-1 PARlated in vitro at multiple glutamate residues within its auto-modification domain, NAD + binding domain and the DNA binding domain. 22 Notably, although a variety of antibodies recognizing the PAR moiety exist, no antibodies against individual proteins modified by PARlation have been generated so far.…”

Section: Poly(adp-ribosyl)ation: the Game And The Playersmentioning

confidence: 99%

Poly(ADP-ribosyl)ation and Epigenetics: Is CTCF PARt of the Plot?

Klenova

Ohlsson²

2004

Cell Cycle

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Enzymological properties of poly(ADP-ribose)polymerase: characterization of automodification sites and NADase activity

Cited by 92 publications

References 0 publications

PARP-1 binds E2F-1 independently of its DNA binding and catalytic domains, and acts as a novel coactivator of E2F-1-mediated transcription during re-entry of quiescent cells into S phase

PARP-1 binds E2F-1 independently of its DNA binding and catalytic domains, and acts as a novel coactivator of E2F-1-mediated transcription during re-entry of quiescent cells into S phase

Signaling Mechanism of Poly(ADP-Ribose) Polymerase-1 (PARP-1) in Inflammatory Diseases

Poly(ADP-ribosyl)ation and Epigenetics: Is CTCF PARt of the Plot?

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