title: Human DNA polymerase theta possesses 5'-dRP lyase activity and functions in single-nucleotide base excision repair in vitro.
abstract: DNA polymerase theta (Pol theta) 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 theta 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 theta 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 theta is a approximately 300-kDa polypeptide, but we show here that the 98-kDa C-terminal region of Pol theta 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 NaBH(4) cross-linking with a BER intermediate revealed that the dRP lyase active site resides in a 24-kDa domain of Pol theta. These results are consistent with a role of Pol theta in BER.
authors.affiliation: Laboratory of Structural Biology, NIEHS, National Institutes of Health, Research Triangle Park, NC 27709, USA.
authors.forename: Rajendra
authors.initials: R
authors.lastname: Prasad
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Longley
authors.forename: Farida S
authors.initials: FS
authors.lastname: Sharief
authors.forename: Esther W
authors.initials: EW
authors.lastname: Hou
authors.forename: William C
authors.initials: WC
authors.lastname: Copeland
authors.forename: Samuel H
authors.initials: SH
authors.lastname: Wilson
mesh_terms: D020134:Catalytic Domain; D004260:DNA Repair; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D010455:Peptides / Q000737:chemistry; D019761:Phosphorus-Oxygen Lyases / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D000097462:DNA Polymerase theta
chemical_list: D004259:DNA-Directed DNA Polymerase; D010455:Peptides; D019761:Phosphorus-Oxygen Lyases; D000097462:DNA Polymerase theta; C423906:5'-deoxyribose phosphate lyase