Three DNA polymerases, recruited by different mechanisms, carry out NER repair synthesis in human cells

Mol Cell. 2010 Mar 12;37(5):714-27. doi: 10.1016/j.molcel.2010.02.009.

Abstract

Nucleotide excision repair (NER) is the most versatile DNA repair system that deals with the major UV photoproducts in DNA, as well as many other DNA adducts. The early steps of NER are well understood, whereas the later steps of repair synthesis and ligation are not. In particular, which polymerases are definitely involved in repair synthesis and how they are recruited to the damaged sites has not yet been established. We report that, in human fibroblasts, approximately half of the repair synthesis requires both pol kappa and pol delta, and both polymerases can be recovered in the same repair complexes. Pol kappa is recruited to repair sites by ubiquitinated PCNA and XRCC1 and pol delta by the classical replication factor complex RFC1-RFC, together with a polymerase accessory factor, p66, and unmodified PCNA. The remaining repair synthesis is dependent on pol epsilon, recruitment of which is dependent on the alternative clamp loader CTF18-RFC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATPases Associated with Diverse Cellular Activities
  • Carrier Proteins / metabolism
  • Cell Line
  • Cellular Senescence
  • DNA Damage*
  • DNA Polymerase II / metabolism
  • DNA Polymerase III / metabolism
  • DNA Repair*
  • DNA-Binding Proteins / metabolism
  • DNA-Directed DNA Polymerase / genetics
  • DNA-Directed DNA Polymerase / metabolism*
  • Fibroblasts / enzymology*
  • Fibroblasts / radiation effects
  • Humans
  • Nuclear Proteins / metabolism
  • Poly-ADP-Ribose Binding Proteins
  • Proliferating Cell Nuclear Antigen / metabolism
  • Protein Processing, Post-Translational
  • Protein Transport
  • RNA Interference
  • Recombinant Fusion Proteins / metabolism
  • Replication Protein C / metabolism
  • Time Factors
  • Transfection
  • Ubiquitin-Protein Ligases
  • Ubiquitination
  • Ultraviolet Rays
  • X-ray Repair Cross Complementing Protein 1

Substances

  • CHTF18 protein, human
  • Carrier Proteins
  • DNA-Binding Proteins
  • Nuclear Proteins
  • Poly-ADP-Ribose Binding Proteins
  • Proliferating Cell Nuclear Antigen
  • RAD18 protein, human
  • RFC1 protein, human
  • Recombinant Fusion Proteins
  • X-ray Repair Cross Complementing Protein 1
  • XRCC1 protein, human
  • Ubiquitin-Protein Ligases
  • POLD1 protein, human
  • POLD3 protein, human
  • DNA Polymerase II
  • DNA Polymerase III
  • DNA-Directed DNA Polymerase
  • POLE protein, human
  • POLK protein, human
  • ATPases Associated with Diverse Cellular Activities
  • Replication Protein C