The noncatalytic C-terminus of AtPOLK Y-family DNA polymerase affects synthesis fidelity, mismatch extension and translesion replication.
Cell survival depends not only on the ability to repair damaged DNA but also on the capability to perform DNA replication on unrepaired or imperfect templates. Crucial to this process are specialized DNA polymerases belonging to the Y family. These enzymes share a similar catalytic fold in their N-terminal region, and most of them have a less-well-conserved C-terminus which is not required for catalytic activity. Although this region is essential for appropriate localization and recruitment in vivo, its precise role during DNA synthesis remains unclear. Here we have compared the catalytic properties of AtPOLK, an Arabidopsis orthologue of mammalian pol kappa, and a truncated version lacking 193 amino acids from its C-terminus. We found that C-terminally truncated AtPOLK is a high-efficiency mutant protein, the DNA-binding capacity of which is not affected but it has higher catalytic efficiency and fidelity than the full-length enzyme. The truncated protein shows increased propensity to extend mispaired primer termini through misalignment and enhanced error-free bypass activity on DNA templates containing 7,8-dihydro-8-oxoGuanine. These results suggest that, in addition to facilitating recruitment to the replication fork, the C-terminus of Y-family DNA polymerases may also play a role in the kinetic control of their enzymatic activity.
García-Ortiz
María Victoria
MV
Departamento de Genética, Universidad de Córdoba, Spain.
Roldán-Arjona
Teresa
T
Ariza
Rafael R
RR
D020816:Amino Acid Motifs; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D020137:Base Pair Mismatch; D001483:Base Sequence; D004247:DNA / Q000737:chemistry; D004249:DNA Damage; D017931:DNA Primers / Q000737:chemistry; D004260:DNA Repair; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000502:physiology*; D007700:Kinetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D017434:Protein Structure, Tertiary
D017931:DNA Primers; D004247:DNA; D004259:DNA-Directed DNA Polymerase