title: Identification of an intrinsic 5'-deoxyribose-5-phosphate lyase activity in human DNA polymerase lambda: a possible role in base excision repair.
abstract: Base excision repair (BER) is a major repair pathway in eukaryotic cells responsible for repair of lesions that give rise to abasic (AP) sites in DNA. Pivotal to this process is the 5'-deoxyribose-5-phosphate lyase (dRP lyase) activity of DNA polymerase beta (Pol beta). DNA polymerase lambda (Pol lambda) is a recently identified eukaryotic DNA polymerase that is homologous to Pol beta. We show here that human Pol lambda exhibits dRP lyase, but not AP lyase, activity in vitro and that this activity is consistent with a beta-elimination mechanism. Accordingly, a single amino acid substitution (K310A) eliminated more than 90% of the wild-type dRP lyase activity, thus suggesting that Lys(310) of Pol lambda is the main nucleophile involved in the reaction. The dRP lyase activity of Pol lambda, in coordination with its polymerization activity, efficiently repaired uracil-containing DNA in an in vitro reconstituted BER reaction. These results suggest that Pol lambda may participate in "single-nucleotide" base excision repair in mammalian cells.
authors.affiliation: Centro de Biologia Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma, 28049 Madrid, Spain.
authors.forename: M
authors.initials: M
authors.lastname: García-Díaz
authors.forename: K
authors.initials: K
authors.lastname: Bebenek
authors.forename: T A
authors.initials: TA
authors.lastname: Kunkel
authors.forename: L
authors.initials: L
authors.lastname: Blanco
mesh_terms: D004260:DNA Repair; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D019761:Phosphorus-Oxygen Lyases / Q000032:analysis / Q000502:physiology*; D013329:Structure-Activity Relationship
chemical_list: C423906:5'-deoxyribose phosphate lyase; C109534:DNA polymerase X; D004259:DNA-Directed DNA Polymerase; D019761:Phosphorus-Oxygen Lyases