Article Dans Une Revue (Data Paper) Methods in Molecular Biology Année : 2024

Studying Translesion DNA Synthesis Using Xenopus In Vitro Systems

Antoine Aze
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James Hutchins
Domenico Maiorano

Résumé

Cell-free extracts derived from Xenopus eggs have been widely used to decipher molecular pathways involved in several cellular processes including DNA synthesis, the DNA damage response, and genome integrity maintenance. We set out assays using Xenopus cell-free extracts to study translesion DNA synthesis (TLS), a branch of the DNA damage tolerance pathway that allows replication of damaged DNA. Using this system, we were able to recapitulate TLS activities that occur naturally in vivo during early embryogenesis. This chapter describes protocols to detect chromatin-bound TLS factors by western blotting and immunofluorescence microscopy upon induction of DNA damage by UV irradiation, monitor TLS-dependent mutagenesis, and perform proteomic screening.

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Dates et versions

inserm-04620024 , version 1 (21-06-2024)

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Antoine Aze, James Hutchins, Domenico Maiorano. Studying Translesion DNA Synthesis Using Xenopus In Vitro Systems. Methods in Molecular Biology, 2024, Methods in Molecular Biology, 2740, pp.21-36. ⟨10.1007/978-1-0716-3557-5_2⟩. ⟨inserm-04620024⟩
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