title: Identification of candidate substrates for poly(ADP-ribose) polymerase-2 (PARP2) in the absence of DNA damage using high-density protein microarrays.
abstract: Poly(ADP-ribose) polymerase-2 (PARP2) belongs to the ADP-ribosyltransferase family of enzymes that catalyze the addition of ADP-ribose units to acceptor proteins, thus affecting many diverse cellular processes. In particular, PARP2 shares with PARP1 and, as recently highlighted, PARP3 the sole property of being catalytically activated by DNA-strand breaks, implying key downstream functions in the cellular response to DNA damage for both enzymes. However, evidence from several studies suggests unique functions for PARP2 in additional processes, possibly mediated through its basal, DNA-damage unstimulated ADP-ribosylating activity. Here, we describe the development and application of a protein microarray-based approach tailored to identify proteins that are ADP-ribosylated by PARP2 in the absence of DNA damage mimetics and might thus represent useful entry points to the exploration of novel PARP2 functions. Several candidate substrates for PARP2 were identified and global hit enrichment analysis showed a clear enrichment in translation initiation and RNA helicase molecular functions. In addition, the top scoring candidates FK506-binding protein 3 and SH3 and cysteine-rich domain-containing protein 1 were selected and confirmed in a complementary assay format as substrates for unstimulated PARP2.
authors.affiliation: Department of Biotechnology, BU Oncology, Nerviano Medical Sciences Srl, Nerviano (MI), Italy.
authors.forename: Sonia
authors.initials: S
authors.lastname: Troiani
authors.forename: Rosita
authors.initials: R
authors.lastname: Lupi
authors.forename: Rita
authors.initials: R
authors.lastname: Perego
authors.forename: Stefania Re
authors.initials: SR
authors.lastname: Depaolini
authors.forename: Sandrine
authors.initials: S
authors.lastname: Thieffine
authors.forename: Roberta
authors.initials: R
authors.lastname: Bosotti
authors.forename: Luisa
authors.initials: L
authors.lastname: Rusconi
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D006801:Humans; D064111:Minichromosome Maintenance Complex Component 2; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods*; D022021:Tacrolimus Binding Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; C074856:FKBP3 protein, human; C103302:STAC protein, human; D000244:Adenosine Diphosphate; C483041:PARP2 protein, human; D011065:Poly(ADP-ribose) Polymerases; C084859:MCM2 protein, human; D064111:Minichromosome Maintenance Complex Component 2; D022021:Tacrolimus Binding Proteins