DNA sequence context greatly affects the accuracy of bypass across an ultraviolet light 6-4 photoproduct in mammalian cells

Pola Shriber; Yael Leitner-Dagan; Nicholas Geacintov; Tamar Paz-Elizur; Zvi Livneh

doi:10.1016/j.mrfmmm.2015.08.002

DNA sequence context greatly affects the accuracy of bypass across an ultraviolet light 6-4 photoproduct in mammalian cells

Pola Shriber, Yael Leitner-Dagan, Nicholas Geacintov, Tamar Paz-Elizur, Zvi Livneh

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism carried out by low-fidelity DNA polymerases that bypass DNA lesions, which overcomes replication stalling. Despite the miscoding nature of most common DNA lesions, several of them are bypassed in mammalian cells in a relatively accurate manner, which plays a key role maintaining a low mutation load. Whereas it is generally agreed that TLS across the major UV and sunlight induced DNA lesion, the cyclobutane pyrimidine dimer (CPD), is accurate, there were conflicting reports on whether the same is true for the thymine-thymine pyrimidine-pyrimidone(6-4) ultraviolet light photoproduct (TT6-4PP), which represents the second most common class of UV lesions. Using a TLS assay system based on gapped plasmids carrying site-specific TT6-4PP lesions in defined sequence contexts we show that the DNA sequence context markedly affected both the extent and accuracy of TLS. The sequence exhibiting higher TLS exhibited also higher error-frequency, caused primarily by semi-targeted mutations, at the nearest nucleotides flanking the lesion. Our results resolve the discrepancy reported on TLS across TT6-4PP, and suggest that TLS is more accurate in human cells than in mouse cells.

Original language	English
Pages (from-to)	71-76
Number of pages	6
Journal	Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
Volume	780
DOIs	https://doi.org/10.1016/j.mrfmmm.2015.08.002
State	Published - 1 Oct 2015
Externally published	Yes

Bibliographical note

Publisher Copyright:
© 2015 Elsevier B.V.

Funding

We thank Alan Lehmann (University of Sussex, Brighton, U.K) and Niels de Wind (Department of Toxicogenetics, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, The Netherlands) for providing cell lines. Z.L. is the incumbent of the Maxwell Ellis Chair for Biomedical Research. This work was supported by grants from the Flight Attendant Medical Research Institute , Florida, USA (to ZL and TPE); the Israel Science Foundation Grant 684/12 to ZL; the Leona M. and Harry B. Helmsley Charitable Trust , NY, USA (to ZL). The preparation of the site-specifically modified oligonucleotides was supported by NIH grant CA168469 to N.E.G.

Funders	Funder number
National Institutes of Health
National Cancer Institute	R01CA168469
Flight Attendant Medical Research Institute
Leona M. and Harry B. Helmsley Charitable Trust
Israel Science Foundation	684/12

Keywords

DNA damage
DNA repair
Error-prone repair
Lesion bypass
UV mutagenesis

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10.1016/j.mrfmmm.2015.08.002

Cite this

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title = "DNA sequence context greatly affects the accuracy of bypass across an ultraviolet light 6-4 photoproduct in mammalian cells",

abstract = "Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism carried out by low-fidelity DNA polymerases that bypass DNA lesions, which overcomes replication stalling. Despite the miscoding nature of most common DNA lesions, several of them are bypassed in mammalian cells in a relatively accurate manner, which plays a key role maintaining a low mutation load. Whereas it is generally agreed that TLS across the major UV and sunlight induced DNA lesion, the cyclobutane pyrimidine dimer (CPD), is accurate, there were conflicting reports on whether the same is true for the thymine-thymine pyrimidine-pyrimidone(6-4) ultraviolet light photoproduct (TT6-4PP), which represents the second most common class of UV lesions. Using a TLS assay system based on gapped plasmids carrying site-specific TT6-4PP lesions in defined sequence contexts we show that the DNA sequence context markedly affected both the extent and accuracy of TLS. The sequence exhibiting higher TLS exhibited also higher error-frequency, caused primarily by semi-targeted mutations, at the nearest nucleotides flanking the lesion. Our results resolve the discrepancy reported on TLS across TT6-4PP, and suggest that TLS is more accurate in human cells than in mouse cells.",

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AU - Livneh, Zvi

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AB - Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism carried out by low-fidelity DNA polymerases that bypass DNA lesions, which overcomes replication stalling. Despite the miscoding nature of most common DNA lesions, several of them are bypassed in mammalian cells in a relatively accurate manner, which plays a key role maintaining a low mutation load. Whereas it is generally agreed that TLS across the major UV and sunlight induced DNA lesion, the cyclobutane pyrimidine dimer (CPD), is accurate, there were conflicting reports on whether the same is true for the thymine-thymine pyrimidine-pyrimidone(6-4) ultraviolet light photoproduct (TT6-4PP), which represents the second most common class of UV lesions. Using a TLS assay system based on gapped plasmids carrying site-specific TT6-4PP lesions in defined sequence contexts we show that the DNA sequence context markedly affected both the extent and accuracy of TLS. The sequence exhibiting higher TLS exhibited also higher error-frequency, caused primarily by semi-targeted mutations, at the nearest nucleotides flanking the lesion. Our results resolve the discrepancy reported on TLS across TT6-4PP, and suggest that TLS is more accurate in human cells than in mouse cells.

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DNA sequence context greatly affects the accuracy of bypass across an ultraviolet light 6-4 photoproduct in mammalian cells

Abstract

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Keywords

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