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A complex suite of loci and elements in eukaryotic type II topoisomerases determine selective sensitivity to distinct poisoning agents.

Lookup NU author(s): Professor Caroline Austin

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

Type II topoisomerases catalyze essential DNA transactions and are proven drug targets. Drug discrimination by prokaryotic and eukaryotic topoisomerases is vital to therapeutic utility, but is poorly understood. We developed a next-generation sequencing (NGS) approach to identify drug-resistance mutations in eukaryotic topoisomerases. We show that alterations conferring resistance to poisons of human and yeast topoisomerase II derive from a rich mutational ‘landscape’ of amino acid substitutions broadly distributed throughout the entire enzyme. Both general and discriminatory drug-resistant behaviors are found to arise from different point mutations found at the same amino acid position and to occur far outside known drug-binding sites. Studies of selected resistant enzymes confirm the NGS data and further show that the anti-cancer quinolone vosaroxin acts solely as an intercalating poison, and that the antibacterial ciprofloxacin can poison yeast topoisomerase II. The innate drug-sensitivity of the DNA binding and cleavage region of human and yeast topoisomerases (particularly hTOP2β) is additionally revealed to be significantly regulated by the enzymes’ adenosine triphosphatase regions. Collectively, these studies highlight the utility of using NGS-based methods to rapidly map drug resistance landscapes and reveal that the nucleotide turnover elements of type II topoisomerases impact drug specificity.


Publication metadata

Author(s): Blower TR, Bandak A, Lee ASY, Austin CA, Nitiss JL, Berger JM

Publication type: Article

Publication status: Published

Journal: Nucleic Acid Research

Year: 2019

Volume: 47

Issue: 15

Pages: 8163-8179

Online publication date: 09/07/2019

Acceptance date: 03/07/2019

Date deposited: 03/10/2019

ISSN (print): 1362-4962

ISSN (electronic): 0305-1048

Publisher: Oxford University Press

URL: https://doi.org/10.1093/nar/gkz579


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