[논문]A switch between DNA polymerases δ and λ promotes error-free bypass of 8-oxo-G lesions

Markkanen, Enni; Castrec, Benoî; t; Villani, Giuseppe; Hü; bscher, Ulrich

doi:10.1073/pnas.1211532109

A switch between DNA polymerases δ and λ promotes error-free bypass of 8-oxo-G lesions 원문보기

Proceedings of the National Academy of Sciences of the United States of America, v.109 no.50, 2012년, pp.20401 - 20406

Markkanen, Enni (Institute of Veterinary Biochemistry and Molecular Biology, University of Zü) , Castrec, Benoît (rich-Irchel, 8057 Zurich, Switzerland) , Villani, Giuseppe (and) , Hübscher, Ulrich (Institute of Veterinary Biochemistry and Molecular Biology, University of Zü)

Abstract ▼ AI-Helper

7,8-Dihydro-8-oxoguanine (8-oxo-G) is a highly abundant and mutagenic lesion. Replicative DNA polymerases (pols) are slowed down at 8-oxo-G and insert both correct cytosine (C) and incorrect adenine (A) opposite 8-oxo-G, but they preferentially extend A:8-oxo-G mispairs. Nevertheless, 8-oxo-G bypass is fairly accurate in vivo. Thus, the question how correct bypass of 8-oxo-G lesions is accomplished despite the poor extension of C:8-oxo-G base pairs by replicative pols remains unanswered. Here we show that replicative pol δ pauses in front of 8-oxo-G and displays difficulties extending from correct C:8-oxo-G in contrast to extension from incorrect A:8-oxo-G. This leads to stalling of pol δ at 8-oxo-G after incorporation of correct C. This stalling at C:8-oxo-G can be overcome by a switch from pol δ to pols λ, β, or η, all of which are able to assist pol δ in 8-oxo-G bypass by translesion synthesis (TLS). Importantly, however, only pol λ selectively catalyzes the correct TLS past 8-oxo-G, whereas pols β and η show no selectivity and even preferentially enhance incorrect TLS. The selectivity of pol λ to promote the correct bypass depends on its N-terminal domain. Furthermore, pol λ −/− mouse embryonic fibroblast extracts display reduced 8-oxo-G TLS. Finally, the correct bypass of 8-oxo-G in gapped plasmids in mouse embryonic fibroblasts and HeLa cells is promoted in the presence of pol λ. Our findings suggest that even though 8-oxo-G is not a blocking lesion per se, correct replication over 8-oxo-G is promoted by a pol switch between pols δ and λ.

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A switch between DNA polymerases δ and λ promotes error-free bypass of 8-oxo-G lesions 원문보기

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