Unrepaired base excision repair intermediates in template DNA strands trigger replication fork collapse and PARP inhibitor sensitivity

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Serrano-Benitez, Almudena ; Wells, Sophie E. ; Drummond-Clarke, Lylah ; Russo, Lilian C. ; Thomas, John Christopher ; Leal, Giovanna A. ; Farrow, Mark ; Edgerton, James Michael ; Balasubramanian, Shankar ; Yang, Ming ; Frezza, Christian ; Gautam, Amit ; Brazina, Jan ; Burdova, Kamila ; Hoch, Nicolas C. ; Jackson, Stephen P. ; Caldecott, Keith W.

• Tipo de documento: Artigo Científico
• Fonte: EMBO Journal; v. 42, n. 18, p. 19-pg., 2023-07-26.

Resumo

DNA single-strand breaks (SSBs) disrupt DNA replication and induce chromosome breakage. However, whether SSBs induce chromosome breakage when present behind replication forks or ahead of replication forks is unclear. To address this question, we exploited an exquisite sensitivity of SSB repair-defective human cells lacking PARP activity or XRCC1 to the thymidine analogue 5-chloro-2 & PRIME;-deoxyuridine (CldU). We show that incubation with CldU in these cells results in chromosome breakage, sister chromatid exchange, and cytotoxicity by a mechanism that depends on the S phase activity of uracil DNA glycosylase (UNG). Importantly, we show that CldU incorporation in one cell cycle is cytotoxic only during the following cell cycle, when it is present in template DNA. In agreement with this, while UNG induces SSBs both in nascent strands behind replication forks and in template strands ahead of replication forks, only the latter trigger fork collapse and chromosome breakage. Finally, we show that BRCA-defective cells are hypersensitive to CldU, either alone and/or in combination with PARP inhibitor, suggesting that CldU may have clinical utility. (AU)

• Processo FAPESP: 18/18007-5 - ADP-ribosilação de proteínas: sinalização de danos ao DNA e impactos na saúde humana
• Beneficiário: Nicolas Carlos Hoch
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 19/06039-2 - EMU concedido no processo 2018/18007-5: microscópio TissueFAXS
• Beneficiário: Nicolas Carlos Hoch
• Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários