Human DNA polymerase θ possesses 5′-dRP lyase activity and functions in single-nucleotide base excision repair in vitro

• Published in: Nucleic acids research 2009-04, Vol.37 (6), p.1868-1877
• Main Authors: Prasad, Rajendra, Longley, Matthew J., Sharief, Farida S., Hou, Esther W., Copeland, William C., Wilson, Samuel H.
• Format: Article
• Language: English
• Subjects: Catalytic Domain; DNA Polymerase theta; DNA Repair; DNA-Directed DNA Polymerase - chemistry; DNA-Directed DNA Polymerase - metabolism; Humans; Kinetics; Nucleic Acid Enzymes; Peptides - chemistry; Phosphorus-Oxygen Lyases - chemistry; Phosphorus-Oxygen Lyases - metabolism; Protein Structure, Tertiary
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gkp035

DNA polymerase θ (Pol θ) is a low-fidelity DNA polymerase that belongs to the family A polymerases and has been proposed to play a role in somatic hypermutation. Pol θ has the ability to conduct translesion DNA synthesis opposite an AP site or thymine glycol, and it was recently proposed to be involved in base excision repair (BER) of DNA damage. Here, we show that Pol θ has intrinsic 5′-deoxyribose phosphate (5′-dRP) lyase activity that is involved in single-nucleotide base excision DNA repair (SN-BER). Full-length human Pol θ is a ∼300-kDa polypeptide, but we show here that the 98-kDa C-terminal region of Pol θ possesses both DNA polymerase activity and dRP lyase activity and is sufficient to carry out base excision repair in vitro. The 5′-dRP lyase activity is independent of the polymerase activity, in that a polymerase inactive mutant retained full 5′-dRP lyase activity. Domain mapping of the 98-kDa enzyme by limited proteolysis and NaBH4 cross-linking with a BER intermediate revealed that the dRP lyase active site resides in a 24-kDa domain of Pol θ. These results are consistent with a role of Pol θ in BER.