Error-free replicative bypass of thymine glycol by the combined action of DNA polymerases kappa and zeta in human cells

Proc Natl Acad Sci U S A. 2010 Aug 10;107(32):14116-21. doi: 10.1073/pnas.1007795107. Epub 2010 Jul 26.


Thymine glycol (Tg) is the most common DNA lesion of thymine induced by interaction with reactive oxygen species. Because of the addition of hydroxyl groups at C5 and C6 in a Tg lesion, the damaged base loses its aromatic character and becomes nonplanar; consequently, the C5 methyl group protrudes in an axial direction and that prevents the stacking of the 5' base above the Tg lesion. Because Tg presents a severe block to continued synthesis by replicative DNA polymerases, we determine here how human cells manage to replicate through this lesion. Using a duplex plasmid system where bidirectional replication ensues from an origin of replication, we show that translesion synthesis (TLS) makes a prominent contribution to Tg bypass and that it occurs in a predominantly error-free fashion. Also, we provide evidence that Pol kappa and Pol zeta function together in promoting error-free replication through the lesion, and based on structural and biochemical information, we propose a role for Pol kappa at the insertion step and of Pol zeta at the extension step of Tg bypass. We discuss the implications of these observations and suggest that human cells have adapted the TLS machinery to function in a much more error-free fashion than could have been predicted from the intrinsic catalytic efficiencies and fidelities of TLS polymerases.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • DNA Damage
  • DNA Repair*
  • DNA Replication*
  • DNA-Directed DNA Polymerase / metabolism*
  • Humans
  • Thymine / analogs & derivatives*
  • Thymine / metabolism


  • thymine glycol
  • DNA polymerase zeta
  • DNA-Directed DNA Polymerase
  • POLK protein, human
  • Thymine