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Nucleic Acids Research

Nucleic Acids Research

IF: 16.6
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Real-time imaging of drug-induced trapping of cellular topoisomerases and poly(ADP-ribose) polymerase 1 at the single-molecule level

Published:13 October 2023 DOI: 10.1093/nar/gkad735 PMID: 37670571
Yilun Sun,?Jiji Chen,?Yves Pommier

Abstract

Topoisomerases (TOP1, TOP2α, and β) are nuclear enzymes crucial for virtually all aspects of DNA metabolisms. They also are the targets of important anti-tumor chemotherapeutics that act by trapping the otherwise reversible topoisomerase-DNA covalent complex intermediates (TOPccs) that are formed during their catalytic reactions, resulting in long-lived topoisomerase DNA-protein crosslinks (TOP-DPCs) that interfere with DNA transactions. The Poly(ADP-ribose) polymerase (PARP) family protein PARP1 is activated by DNA damage to recruit DNA repair proteins, and PARP inhibitors are another class of commonly used chemotherapeutics, which bind and trap PARP molecules on DNA. To date, the trapping of TOPccs and PARP by their respective inhibitors can only be measured by immune-biochemical methods in cells. Here, we developed an imaging-based approach enabling real-time monitoring of drug-induced trapping of TOPccs and PARP1 in live cells at the single-molecule level. Capitalizing on this approach, we calculated the fraction of self-fluorescence tag-labeled topoisomerases and PARP single-molecules that are trapped by their respective inhibitors in real time. This novel technique should help elucidate the molecular processes that repair TOPcc and PARP trapping and facilitate the development of novel topoisomerase and PARP inhibitor-based therapies.

Substances (4)

Materials
Procduct Name CAS Molecular Formula Supplier Price
PDD 00017273 1945950-21-9 C23H26N6O4S2 67 suppliers $49.00-$4741.00
PDD 00017273 1945950-21-9 C23H26N6O4S2 67 suppliers $49.00-$4741.00
PDD 00017273 1945950-21-9 C23H26N6O4S2 67 suppliers $49.00-$4741.00
PDD 00017273 1945950-21-9 C23H26N6O4S2 67 suppliers $49.00-$4741.00

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