Poly (ADP-ribose) polymerase inhibits DNA replication by human replicative DNA polymerase alpha, delta and epsilon in vitro

FEBS Lett. 1994 Dec 19;356(2-3):261-6. doi: 10.1016/0014-5793(94)01280-6.

Abstract

The influence of poly (ADP-ribose) polymerase (PARP) and poly ADP-ribosylation on DNA synthesis supported by human replicative DNA polymerase (DNA pol) alpha, delta, and epsilon has been examined using the replication system containing poly(dA)4500-oligo(dT)12-18 as the template primer. PARP alone inhibited the pol activities in a dose-dependent manner even in the presence of the accessory factors for DNA pol delta, proliferating cell nuclear antigen (PCNA) and activator 1 (Al; RF-C). Both DNA pol alpha and epsilon activities were decreased approximately 10-fold under the poly ADP-ribosylating condition. In contrast, DNA synthesis by DNA pol delta holoenzyme was not affected by poly ADP-ribosylation like prokaryotic DNA pol's. The analysis of poly(dT) formed by DNA pol alpha and epsilon indicated that poly ADP-ribosylation mainly reduced the frequency of replication. These observations suggest a possibility that PARP acts as a negative regulator for the initiation of DNA replication upon cellular DNA damage.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Polymerase II / metabolism*
  • DNA Polymerase III
  • DNA Replication*
  • DNA-Directed DNA Polymerase / metabolism*
  • Dose-Response Relationship, Drug
  • HeLa Cells
  • Humans
  • Kinetics
  • Poly Adenosine Diphosphate Ribose / biosynthesis
  • Poly(ADP-ribose) Polymerases / isolation & purification
  • Poly(ADP-ribose) Polymerases / metabolism*
  • Poly(ADP-ribose) Polymerases / pharmacology
  • Templates, Genetic

Substances

  • Poly Adenosine Diphosphate Ribose
  • Poly(ADP-ribose) Polymerases
  • DNA Polymerase II
  • DNA Polymerase III
  • DNA-Directed DNA Polymerase