Co-localization in replication foci and interaction of human Y-family members, DNA polymerase pol eta and REVl protein

DNA Repair (Amst). 2004 Nov 2;3(11):1503-14. doi: 10.1016/j.dnarep.2004.06.015.

Abstract

The progress of replicative DNA polymerases along the replication fork may be impeded by the presence of lesions in the genome. One way to circumvent such hurdles involves the recruitment of specialized DNA polymerases that perform limited incorporation of nucleotides in the vicinity of the damaged site. This process entails DNA polymerase switch between replicative and specialized DNA polymerases. Five eukaryotic proteins can carry out translesion synthesis (TLS) of damaged DNA in vitro, DNA polymerases zeta, eta, iota, and kappa, and REV1. To identify novel proteins that interact with hpol eta, we performed a yeast two-hybrid screen. In this paper, we show that hREV1 interacts with hpol eta as well as with hpol kappa and poorly with hpol iota. Furthermore, cellular localization analysis demonstrates that hREV1 is present, with hpol eta in replication factories at stalled replication forks and is tightly associated with nuclear structures. This hREV1 nuclear localization occurs independently of the presence of hpol eta. Taken together, our data suggest a central role for hREV1 as a scaffold that recruits DNA polymerases involved in TLS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cell Line, Transformed
  • Cell Nucleus / metabolism
  • DNA / biosynthesis
  • DNA / genetics
  • DNA Damage
  • DNA Repair
  • DNA Replication
  • DNA-Directed DNA Polymerase / genetics
  • DNA-Directed DNA Polymerase / metabolism*
  • Humans
  • In Vitro Techniques
  • Nuclear Proteins
  • Nucleotidyltransferases / genetics
  • Nucleotidyltransferases / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • S Phase
  • Two-Hybrid System Techniques
  • Ultraviolet Rays

Substances

  • Nuclear Proteins
  • Recombinant Proteins
  • DNA
  • Nucleotidyltransferases
  • REV1 protein, human
  • DNA-Directed DNA Polymerase
  • Rad30 protein