Functional association of poly(ADP-ribose) polymerase with DNA polymerase alpha-primase complex: a link between DNA strand break detection and DNA replication

Nucleic Acids Res. 1998 Apr 15;26(8):1891-8. doi: 10.1093/nar/26.8.1891.

Abstract

Poly(ADP-ribose) polymerase (PARP) is an element of the DNA damage surveillance network evolved by eukaryotic cells to cope with numerous environmental and endogenous genotoxic agents. PARP has been found to be involved in vivo in both cell proliferation and base excision repair of DNA. In this study the interaction between PARP and the DNA polymerase alpha-primase tetramer has been examined. We provide evidence that in proliferating cells: (i) PARP is physically associated with the catalytic subunit of the DNA polymerase alpha-primase tetramer, an association confirmed by confocal microscopy, demonstrating that both enzymes are co-localized at the nuclear periphery of HeLa cells; (ii) this interaction requires the integrity of the second zinc finger of PARP and is maximal during the S and G2/M phases of the cell cycle; (iii) PARP-deficient cells derived from PARP knock-out mice exhibited reduced DNA polymerase activity, compared with the parental cells, a reduction accentuated following exposure to sublethal doses of methylmethanesulfonate. Altogether, the present results strongly suggest that PARP participates in a DNA damage survey mechanism implying its nick-sensor function as part of the control of replication fork progression when breaks are present in the template.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aphidicolin / pharmacology
  • Binding Sites
  • Cell Cycle / drug effects
  • Cell Cycle / physiology*
  • Cell Division
  • DNA Damage*
  • DNA Polymerase I / chemistry
  • DNA Polymerase I / isolation & purification
  • DNA Polymerase I / metabolism*
  • DNA Primase / chemistry
  • DNA Primase / isolation & purification
  • DNA Primase / metabolism*
  • DNA Repair*
  • DNA Replication*
  • G2 Phase
  • HeLa Cells
  • Humans
  • Kinetics
  • Macromolecular Substances
  • Mice
  • Mice, Knockout
  • Microscopy, Confocal
  • Mitosis
  • Models, Biological
  • Poly(ADP-ribose) Polymerases / chemistry
  • Poly(ADP-ribose) Polymerases / deficiency
  • Poly(ADP-ribose) Polymerases / isolation & purification
  • Poly(ADP-ribose) Polymerases / metabolism*
  • S Phase
  • Templates, Genetic
  • Zinc Fingers

Substances

  • Macromolecular Substances
  • Aphidicolin
  • Poly(ADP-ribose) Polymerases
  • DNA Primase
  • DNA Polymerase I