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

Tim R. Blower, Afif Bandak, Amy S.Y. Lee, Caroline A. Austin, John L. Nitiss, James M Berger

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)8163-8179
Number of pages17
JournalNucleic acids research
Volume47
Issue number15
DOIs
StatePublished - Sep 5 2019

Fingerprint

Type II DNA Topoisomerase
Poisoning
Poisons
Pharmaceutical Preparations
Yeasts
Drug Resistance
Enzymes
DNA Cleavage
Quinolones
Amino Acid Substitution
Ciprofloxacin
Point Mutation
Adenosine Triphosphatases
Nucleotides
Binding Sites
Amino Acids
Mutation
DNA
Neoplasms

ASJC Scopus subject areas

  • Genetics

Cite this

A complex suite of loci and elements in eukaryotic type II topoisomerases determine selective sensitivity to distinct poisoning agents. / Blower, Tim R.; Bandak, Afif; Lee, Amy S.Y.; Austin, Caroline A.; Nitiss, John L.; Berger, James M.

In: Nucleic acids research, Vol. 47, No. 15, 05.09.2019, p. 8163-8179.

Research output: Contribution to journalArticle

Blower, Tim R. ; Bandak, Afif ; Lee, Amy S.Y. ; Austin, Caroline A. ; Nitiss, John L. ; Berger, James M. / A complex suite of loci and elements in eukaryotic type II topoisomerases determine selective sensitivity to distinct poisoning agents. In: Nucleic acids research. 2019 ; Vol. 47, No. 15. pp. 8163-8179.
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