Quantitative measurement of translesion DNA synthesis in mammalian cells

Omer Ziv; Noam Diamant; Sigal Shachar; Ayal Hendel; Zvi Livneh

doi:10.1007/978-1-61779-998-3_35

Quantitative measurement of translesion DNA synthesis in mammalian cells

Omer Ziv
, Noam Diamant
, Sigal Shachar
, Ayal Hendel
, Zvi Livneh

Weizmann Institute of Science

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

11 Scopus citations

Abstract

Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism, in which specialized low-fidelity DNA polymerases bypass lesions that interfere with replication. This process is inherently mutagenic due to the miscoding nature of DNA lesions, but it prevents double strand breaks, genome instability, and cancer. We describe here a quantitative method for measuring TLS in mammalian cells, based on non-replicating plasmids that carry a defined and site-specific DNA lesion in a single-stranded DNA region opposite a gap. The assay is responsive to the cellular composition of TLS DNA polymerases, and TLS regulators. It can be used with a broad variety of cultured mammalian cells, and is amenable to RNAi gene silencing, making it a useful tool in the study of TLS in mammalian cells.

Original language	English
Title of host publication	DNA Repair Protocols
Publisher	Humana Press Inc.
Pages	529-542
Number of pages	14
ISBN (Print)	9781617799976
DOIs	https://doi.org/10.1007/978-1-61779-998-3_35
State	Published - 2012
Externally published	Yes

Publication series

Name	Methods in Molecular Biology
Volume	920
ISSN (Print)	1064-3745

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Carcinogenesis
DNA damage tolerance
DNA repair
Gapped plasmid
Mutagenesis
Site-specific DNA damage
TLS
Translesion DNA Synthesis

Access to Document

10.1007/978-1-61779-998-3_35

Cite this

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keywords = "Carcinogenesis, DNA damage tolerance, DNA repair, Gapped plasmid, Mutagenesis, Site-specific DNA damage, TLS, Translesion DNA Synthesis",

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AU - Ziv, Omer

AU - Diamant, Noam

AU - Shachar, Sigal

AU - Hendel, Ayal

AU - Livneh, Zvi

PY - 2012

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AB - Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism, in which specialized low-fidelity DNA polymerases bypass lesions that interfere with replication. This process is inherently mutagenic due to the miscoding nature of DNA lesions, but it prevents double strand breaks, genome instability, and cancer. We describe here a quantitative method for measuring TLS in mammalian cells, based on non-replicating plasmids that carry a defined and site-specific DNA lesion in a single-stranded DNA region opposite a gap. The assay is responsive to the cellular composition of TLS DNA polymerases, and TLS regulators. It can be used with a broad variety of cultured mammalian cells, and is amenable to RNAi gene silencing, making it a useful tool in the study of TLS in mammalian cells.

KW - Carcinogenesis

KW - DNA damage tolerance

KW - DNA repair

KW - Gapped plasmid

KW - Mutagenesis

KW - Site-specific DNA damage

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KW - Translesion DNA Synthesis

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Quantitative measurement of translesion DNA synthesis in mammalian cells

Abstract

Publication series

UN SDGs

Keywords

Access to Document

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