title: Translesion synthesis across the (6-4) photoproduct and its Dewar valence isomer by the Y-family and engineered DNA polymerases.
abstract: We analyzed the translesion synthesis across the UV-induced lesions, the (6-4) photoproduct and its Dewar valence isomer, by using human DNA polymerases eta and iota in vitro. The primer extension experiments revealed that pol eta tended to incorporate dG opposite the 3' component of both lesions, but the incorporation efficiency for the Dewar isomer was higher than that for the (6-4) photoproduct. On the other hand, pol iota was likely to incorporate dA opposite the 3' components of the (6-4) photoproduct and its Dewar isomer with a similar efficiency. Elongation after the incorporation opposite the UV lesions was not observed for these Y-family polymerases. We further analyzed the bypass ability of an engineered polymerase developed from Thermus DNA polymerase for the amplification of ancient DNA. This polymerase could bypass the Dewar isomer more efficiently than the (6-4) photoproduct.
authors.affiliation: Division of Chemisty, Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka 563-8531, Japan.
authors.forename: Junpei
authors.initials: J
authors.lastname: Yamamoto
authors.forename: David
authors.initials: D
authors.lastname: Loakes
authors.forename: Chikahide
authors.initials: C
authors.lastname: Masutani
authors.forename: Shizu
authors.initials: S
authors.lastname: Simmyo
authors.forename: Kumiko
authors.initials: K
authors.lastname: Urabe
authors.forename: Fumio
authors.initials: F
authors.lastname: Hanaoka
authors.forename: Philipp
authors.initials: P
authors.lastname: Holliger
authors.forename: Shigenori
authors.initials: S
authors.lastname: Iwai
mesh_terms: D004247:DNA / Q000096:biosynthesis / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007536:Isomerism; D009154:Mutation; D015202:Protein Engineering; D011740:Pyrimidine Dimers / Q000737:chemistry*; D013824:Thermus / Q000201:enzymology; D000094769:DNA Polymerase iota; D000099270:Y-Family DNA Polymerases
chemical_list: D004247:DNA; D004259:DNA-Directed DNA Polymerase; D011740:Pyrimidine Dimers; D000094769:DNA Polymerase iota; C058406:pyrimidine-pyrimidone dimer; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases