Detection of endogenous translesion DNA synthesis in single mammalian cells

Egger, Tom; Aze, Antoine; Maiorano, Domenico

doi:10.1016/j.crmeth.2023.100501

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Cited by 3 publications

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“…The protocol below is inspired by a previous method to detect chromatin-bound proteins 2 and describes step-by-step procedures to detect chromatin-bound TLS factors in HCT116 cells. 1 However, we have also used this protocol with HEK293, T98G and H1299 cells. It may require adaptation for other cell lines requesting specific media (such as stem cells for instance).…”

Section: Before You Beginmentioning

confidence: 99%

Protocol to analyze endogenous translesion DNA synthesis in single mammalian cells

2023

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Section: Before You Beginmentioning

confidence: 99%

Protocol to analyze endogenous translesion DNA synthesis in single mammalian cells

2023

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Studying Translesion DNA Synthesis Using Xenopus In Vitro Systems

Aze,

Hutchins,

Maiorano

2024

Methods in Molecular Biology

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Canonical and Non-Canonical Roles of Human DNA Polymerase η

Bedaiwi,

Usmani,

Carty

2024

Genes

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DNA damage tolerance pathways that allow for the completion of replication following fork arrest are critical in maintaining genome stability during cell division. The main DNA damage tolerance pathways include strand switching, replication fork reversal and translesion synthesis (TLS). The TLS pathway is mediated by specialised DNA polymerases that can accommodate altered DNA structures during DNA synthesis, and are important in allowing replication to proceed after fork arrest, preventing fork collapse that can generate more deleterious double-strand breaks in the genome. TLS may occur directly at the fork, or at gaps remaining behind the fork, in the process of post-replication repair. Inactivating mutations in the human POLH gene encoding the Y-family DNA polymerase Pol η causes the skin cancer-prone genetic disease xeroderma pigmentosum variant (XPV). Pol η also contributes to chemoresistance during cancer treatment by bypassing DNA lesions induced by anti-cancer drugs including cisplatin. We review the current understanding of the canonical role of Pol η in translesion synthesis following replication arrest, as well as a number of emerging non-canonical roles of the protein in other aspects of DNA metabolism.

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Detection of endogenous translesion DNA synthesis in single mammalian cells

Cited by 3 publications

References 50 publications

Protocol to analyze endogenous translesion DNA synthesis in single mammalian cells

Protocol to analyze endogenous translesion DNA synthesis in single mammalian cells

Studying Translesion DNA Synthesis Using Xenopus In Vitro Systems

Canonical and Non-Canonical Roles of Human DNA Polymerase η

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