title: Quantitative measurement of translesion DNA synthesis in mammalian cells.
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.
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Omer
authors.initials: O
authors.lastname: Ziv
authors.forename: Noam
authors.initials: N
authors.lastname: Diamant
authors.forename: Sigal
authors.initials: S
authors.lastname: Shachar
authors.forename: Ayal
authors.initials: A
authors.lastname: Hendel
authors.forename: Zvi
authors.initials: Z
authors.lastname: Livneh
mesh_terms: D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002875:Chromosomes / Q000235:genetics; D004249:DNA Damage; D004277:DNA, Single-Stranded / Q000096:biosynthesis* / Q000235:genetics*; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics; D009838:Oligodeoxyribonucleotides / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids / Q000235:genetics
chemical_list: D004277:DNA, Single-Stranded; D009838:Oligodeoxyribonucleotides