Centromere Proteins Cenpa, Cenpb, and Bub3 Interact With poly(ADP-ribose) polymerase-1 Protein and Are poly(ADP-ribosyl)ated

J Biol Chem. 2002 Jul 26;277(30):26921-6. doi: 10.1074/jbc.M200620200. Epub 2002 May 13.

Abstract

Poly(ADP-ribose) polymerase-1 (PARP-1) is activated by DNA strand breaks during cellular genotoxic stress response and catalyzes poly(ADP-ribosyl)ation of acceptor proteins. These acceptor proteins include those involved in modulation of chromatin structure, DNA synthesis, DNA repair, transcription, and cell cycle control. Thus, PARP-1 is believed to play a pivotal role in maintaining genome integrity through modulation of protein-protein and protein-DNA interactions. We previously described the association of PARP-1 with normal mammalian centromeres and human neocentromeres by affinity purification and immunofluorescence. Here we investigated the interaction of this protein with, and poly(ADP-ribosyl)ation of, three constitutive centromere proteins, Cenpa, Cenpb, and Cenpc, and a spindle checkpoint protein, Bub3. Immunoprecipitation and Western blot analyses demonstrate that Cenpa, Cenpb, and Bub3, but not Cenpc, interacted with PARP-1, and are poly(ADP-ribosyl)ated following induction of DNA damage. The results suggest a role of PARP-1 in centromere assembly/disassembly and checkpoint control. Demonstration of PARP-1-binding and poly(ADP-ribosyl)ation in three of the four proteins tested further suggests that many more centromere proteins may behave similarly and implicates PARP-1 as an important regulator of diverse centromere function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Autoantigens*
  • Binding Sites
  • Blotting, Western
  • Cell Cycle Proteins / metabolism*
  • Cell Line
  • Cell Nucleus / metabolism
  • Centromere / metabolism*
  • Centromere Protein A
  • Centromere Protein B
  • Chromosomal Proteins, Non-Histone / metabolism*
  • DNA Repair
  • DNA-Binding Proteins*
  • Mice
  • Molecular Sequence Data
  • Phenotype
  • Phosphorylation
  • Poly Adenosine Diphosphate Ribose / metabolism*
  • Poly(ADP-ribose) Polymerases / metabolism*
  • Poly-ADP-Ribose Binding Proteins
  • Precipitin Tests
  • Protein Binding
  • Transcription, Genetic

Substances

  • Autoantigens
  • BUB3 protein, human
  • Bub3 protein, mouse
  • CENPA protein, human
  • Cell Cycle Proteins
  • Cenpa protein, mouse
  • Cenpb protein, mouse
  • Centromere Protein A
  • Centromere Protein B
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • Poly-ADP-Ribose Binding Proteins
  • centromere protein C
  • Poly Adenosine Diphosphate Ribose
  • Poly(ADP-ribose) Polymerases