Oxidative guanine base damage plays a dual role in regulating productive ALT-associated homology-directed repair.

Cancer cells maintain telomeres by upregulating telomerase or alternative lengthening of telomeres (ALT) via homology-directed repair at telomeric DNA breaks. 8-Oxoguanine (8oxoG) is a highly prevalent endogenous DNA lesion in telomeric sequences, altering telomere structure and telomerase activity, but its impact on ALT is unclear. Here, we demonstrate that targeted 8oxoG formation at telomeres stimulates ALT activity and homologous recombination specifically in ALT cancer cells. Mechanistically, an acute 8oxoG induction increases replication stress, as evidenced by increased telomere fragility and ATR kinase activation at ALT telomeres. Furthermore, ALT cells are more sensitive to chronic telomeric 8oxoG damage than telomerase-positive cancer cells, consistent with increased 8oxoG-induced replication stress. However, telomeric 8oxoG production in G2 phase, when ALT telomere elongation occurs, impairs telomeric DNA synthesis. Our study demonstrates that a common oxidative base lesion has a dual role in regulating ALT depending on when the damage arises in the cell cycle. [Display omitted] • The oxidative DNA lesion 8-oxoguanine impairs DNA replication at ALT telomeres • 8-Oxoguanine-induced replication stress stimulates ALT homology-directed repair activity • ALT cancer cells are hypersensitive to chronic telomeric 8-oxoguanine damage • 8-Oxoguanine formation in G2 cell phase impairs ALT-replisome telomere elongation Thosar et al. show that oxidative base damage 8-oxoguanine regulates ALT-mediated telomere synthesis depending on when the lesion arises during the cell cycle. 8-Oxoguanine stimulates ALT by impairing DNA replication forks at telomeres, thereby provoking homology-directed repair. However, 8-oxoguanine, when produced in G2 phase, inhibits telomere elongation by the ALT replisome. [ABSTRACT FROM AUTHOR]