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Lookup NU author(s): Kathryn Gilroy, Chrysoula Leontiou, Dr Kay Padget, Professor Jeremy LakeyORCiD, Professor Caroline AustinORCiD
Type II Human DNA Topoisomerases (topos II) play an essential role in DNA replication and transcription and are important targets for cancer chemotherapeutic drugs. Topoisomerase II causes transient double-strand breaks in DNA, forming a gate through which another double helix is passed, and acts as a DNA dependent ATPase. Mutations in topoII have been linked to atypical multi-drug resistance. Both human Topoisomerase II isoforms, α and β, are targeted by amsacrine. We have used a forced molecular evolution approach to identify mutations conferring resistance to acridines. Here we report mutation βG465D, which was selected with mAMSA and DACA and is cross-resistant to etoposide, ellipticine and doxorubicin. Resistance to mAMSA appears to decrease over time indicating a previously unreported resistance mechanism. G465D lies within the B′ domain in the region that contacts the cleaved gate helix. There is a 3-fold decrease in ATP affinity and ATP hydrolysis and an altered requirement for magnesium in decatenation assays. The decatenation rate is decreased for the mutated G465D protein. And we report for the first time the use of fluorescence anisotropy with intact human topoisomerase II. © 2006 Oxford University Press.
Author(s): Gilroy KL, Leontiou C, Padget K, Lakey JH, Austin CA
Publication type: Article
Publication status: Published
Journal: Nucleic Acids Research
Year: 2006
Volume: 34
Issue: 5
Pages: 1597-1607
Print publication date: 01/01/2006
ISSN (print): 0305-1048
ISSN (electronic): 1362-4962
Publisher: Oxford University Press
URL: http://dx.doi.org/10.1093/nar/gkl057
DOI: 10.1093/nar/gkl057
PubMed id: 16549872
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