Translesion synthesis across the (6-4) photoproduct and its Dewar valence isomer by the Y-family and engineered DNA polymerases.
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.
Yamamoto
Junpei
J
Division of Chemisty, Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka 563-8531, Japan.
Loakes
David
D
Masutani
Chikahide
C
Simmyo
Shizu
S
Urabe
Kumiko
K
Hanaoka
Fumio
F
Holliger
Philipp
P
Iwai
Shigenori
S
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
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