title: Poly(ADP-ribose) polymerase 2 localizes to mammalian active centromeres and interacts with PARP-1, Cenpa, Cenpb and Bub3, but not Cenpc.
abstract: Poly(ADP-ribose) polymerase 2 (PARP-2) is a newly discovered member of the PARP family. We report the association of PARP-2 with mammalian centromeres in a cell-cycle-dependent manner, accumulating at centromeres during prometaphase and metaphase, disassociating during anaphase, and disappearing from the centromeres by telophase. Analysis of a pseudodicentric chromosome and a human neocentromere indicates that PARP-2 binding occurs only at active centromeres in a sequence-independent manner. Centromere binding peaks at the outer centromere region, and is significantly enhanced upon treatment with microtubule-inhibiting drugs. Co-immunoprecipitation assay demonstrates interaction between PARP-2 and its functional homolog PARP-1, constitutive centromere proteins Cenpa and Cenpb, and spindle checkpoint protein Bub3, but not with a third constitutive centromere protein Cenpc. These results, together with our previous demonstration that PARP-1 displays an identical binding pattern with Cenpa, Cenpb and Bub3, but not Cenpc, and that all three proteins undergo significant poly(ADP-ribosyl)ation upon gamma-irradiation of cells, point to possible diverse roles of PARP-2 and PARP-1 in modulating the structure and checkpoint functions of the mammalian centromere, in particular during radiation-induced DNA damage.
authors.affiliation: The Murdoch Childrens Research Institute, Royal Children's Hospital, Flemington Road, Parkville 3052, Australia.
authors.forename: Alka
authors.initials: A
authors.lastname: Saxena
authors.forename: Lee H
authors.initials: LH
authors.lastname: Wong
authors.forename: Paul
authors.initials: P
authors.lastname: Kalitsis
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Earle
authors.forename: Lisa G
authors.initials: LG
authors.lastname: Shaffer
authors.forename: K H Andy
authors.initials: KH
authors.lastname: Choo
mesh_terms: D000818:Animals; D001324:Autoantigens; D018797:Cell Cycle Proteins / Q000378:metabolism; D002503:Centromere / Q000378:metabolism*; D000076247:Centromere Protein A; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004268:DNA-Binding Proteins; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008938:Mitosis / Q000502:physiology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D000075223:Poly-ADP-Ribose Binding Proteins
chemical_list: D001324:Autoantigens; C489444:BUB3 protein, human; C489445:Bub3 protein, mouse; C000619700:CENPA protein, human; C497926:CENPB protein, human; D018797:Cell Cycle Proteins; C000619701:Cenpa protein, mouse; C497927:Cenpb protein, mouse; D000076247:Centromere Protein A; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; C060905:centromere protein C; D011065:Poly(ADP-ribose) Polymerases