DNA topoisomerases are the targets for a wide range of clinically useful anti-cancer drugs. Although the basic biochemical reaction pathway for the topoisomerases is fairly well understood, there is still little information available regarding the mechanism of action of the anti-topoisomerase drugs in generating the stable covalent complex that is necessary for cell killing. Studies are proposed to carry out a detailed mutational analysis of eukaryotic topoisomerase II, with the goal of identifying sites in the enzyme which interact with complex stabilizing topoisomerase II inhibitors. Yeast is used as a model system for analyzing the effects of mutant proteins, and for overexpression of the mutant proteins. The research strategy involves the construction of novel topoisomerase mutants, including mutants that are hypersensitive to anti-topoisomerase drugs. In addition to etoposide, studies will focus on other agents with different types of interaction with topoisomerase II, such as ICRF-187, which is a catalytic inhibitor of the enzyme. Mutations will also be introduced into the human topoisomerase II alpha and be at genes to verify that the effects of mutations are similar among the yeast enzymes and the human proteins. A new system is being developed to investigate whether mutations in topoisomerase play a role in the development of acquired drug resistance. Recent findings indicating that topoisomerase I may be an important determinant in cell killing by other DNA damaging agents have led to studies of how cells may modulate topoisomerases following DNA damage. This work is designed to provide insight into the biochemical mechanisms of anti-topoisomerase drug action, and may be useful in the development of new and more effective topoisomerase inhibitors.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA052814-10
Application #
2894850
Study Section
Experimental Therapeutics Subcommittee 1 (ET)
Program Officer
Wolpert, Mary K
Project Start
1997-06-01
Project End
2000-05-31
Budget Start
1999-06-01
Budget End
2000-05-31
Support Year
10
Fiscal Year
1999
Total Cost
Indirect Cost
Name
St. Jude Children's Research Hospital
Department
Type
DUNS #
067717892
City
Memphis
State
TN
Country
United States
Zip Code
38105
Tombline, Gregory; Millen, Jonathan I; Polevoda, Bogdan et al. (2017) Effects of an unusual poison identify a lifespan role for Topoisomerase 2 in Saccharomyces cerevisiae. Aging (Albany NY) 9:68-97
Heo, Jinho; Li, Jing; Summerlin, Matthew et al. (2015) TDP1 promotes assembly of non-homologous end joining protein complexes on DNA. DNA Repair (Amst) 30:28-37
Katyal, Sachin; Lee, Youngsoo; Nitiss, Karin C et al. (2014) Aberrant topoisomerase-1 DNA lesions are pathogenic in neurodegenerative genome instability syndromes. Nat Neurosci 17:813-21
Nitiss, Karin C; Nitiss, John L (2014) Twisting and ironing: doxorubicin cardiotoxicity by mitochondrial DNA damage. Clin Cancer Res 20:4737-9
Gao, Rui; Schellenberg, Matthew J; Huang, Shar-Yin N et al. (2014) Proteolytic degradation of topoisomerase II (Top2) enables the processing of Top2┬ĚDNA and Top2┬ĚRNA covalent complexes by tyrosyl-DNA-phosphodiesterase 2 (TDP2). J Biol Chem 289:17960-9
Nitiss, John L; Nitiss, Karin C (2013) Tdp2: a means to fixing the ends. PLoS Genet 9:e1003370
Hasinoff, Brian B; Wu, Xing; Nitiss, John L et al. (2012) The anticancer multi-kinase inhibitor dovitinib also targets topoisomerase I and topoisomerase II. Biochem Pharmacol 84:1617-26
Nitiss, John L; Soans, Eroica; Rogojina, Anna et al. (2012) Topoisomerase assays. Curr Protoc Pharmacol Chapter 3:Unit 3.3.
Bahmed, Karim; Nitiss, Karin C; Nitiss, John L (2010) Yeast Tdp1 regulates the fidelity of nonhomologous end joining. Proc Natl Acad Sci U S A 107:4057-62
Nitiss, John L (2009) Targeting DNA topoisomerase II in cancer chemotherapy. Nat Rev Cancer 9:338-50

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