Cancer Research The Future of Cancer Research: Science and Patient Impact Cancer Health Disparities Conference 2009
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online
[Cancer Research 59, 3442-3450, July 1, 1999]
© 1999 American Association for Cancer Research
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wessel, I.
Right arrow Articles by Sehested, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wessel, I.
Right arrow Articles by Sehested, M.
[Cancer Research 59, 3442-3450, July 15, 1999]
© 1999 American Association for Cancer Research

Experimental Therapeutics

Human Small Cell Lung Cancer NYH Cells Selected for Resistance to the Bisdioxopiperazine Topoisomerase II Catalytic Inhibitor ICRF-187 Demonstrate a Functional R162Q Mutation in the Walker A Consensus ATP Binding Domain of the {alpha} Isoform1

Irene Wessel, Lars H. Jensen, Peter B. Jensen, Jacob Falck, Angela Rose, Mikael Roerth, John L. Nitiss and Maxwell Sehested2

Department of Pathology, Laboratory Center, Rigshospitalet 5444 [I. W., J. F., M. S.], and Laboratory for Experimental Medical Oncology, Finsen Center, Rigshospitalet 5074 [I. W., L. H. J., P. B. J., J. F., M. R.], DK-2100 Copenhagen, Denmark; and Department of Molecular Pharmacology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101 [L. H. J., A. R., J. L. N.]

Bisdioxopiperazine drugs such as ICRF-187 are catalytic inhibitors of DNA topoisomerase II, with at least two effects on the enzyme: namely, locking it in a closed-clamp form and inhibiting its ATPase activity. This is in contrast to topoisomerase II poisons as etoposide and amsacrine (m-AMSA), which act by stabilizing enzyme-DNA-drug complexes at a stage in which the DNA gate strand is cleaved and the protein is covalently attached to DNA. Human small cell lung cancer NYH cells selected for resistance to ICRF-187 (NYH/187) showed a 25% increase in topoisomerase II{alpha} level and no change in expression of the ß isoform. Sequencing of the entire topoisomerase II{alpha} cDNA from NYH/187 cells demonstrated a homozygous G->A point mutation at nucleotide 485, leading to a R162Q conversion in the Walker A consensus ATP binding site (residues 161–165 in the {alpha} isoform), this being the first drug-selected mutation described at this site. Western blotting after incubation with ICRF-187 showed no depletion of the {alpha} isoform in NYH/187 cells in contrast to wild-type (wt) cells, whereas equal depletion of the ß isoform was observed in the two sublines. Alkaline elution assay demonstrated a lack of inhibition of etoposide-induced DNA single-stranded breaks in NYH/187 cells, whereas this inhibition was readily apparent in NYH cells. Site-directed mutagenesis in human topoisomerase II{alpha} introduced into a yeast Saccharomyces cerevisiae strain with a temperature-conditional yeast TOP2 mutant demonstrated that R162Q conferred resistance to the bisdioxopiperazines ICRF-187 and -193 but not to etoposide or m-AMSA. Both etoposide and m-AMSA induced more DNA cleavage with purified R162Q enzyme than with the wt. The R162Q enzyme has a 20–25% decreased catalytic capacity compared to the wt and was almost inactive at <0.25 mM ATP compared to the wt. Kinetoplast DNA decatenation by the R162Q enzyme at 1 mM ATP was not resistant to ICRF-187 compared to wt, whereas it was clearly less sensitive than wt to ICRF-187 at low ATP concentrations. This suggests that it is a shift in the equilibrium to an open-clamp state in the enzyme’s catalytic cycle caused by a decreased ATP binding by the mutated enzyme that is responsible for bisdioxopiperazine resistance.




This article has been cited by other articles:


Home page
 Mol. Pharmacol.Home page
M. Grauslund, A. V. Thougaard, A. Fuchtbauer, K. F. Hofland, P. H. Hjorth, P. B. Jensen, M. Sehested, E.-M. Fuchtbauer, and L. H. Jensen
A Mouse Model for Studying the Interaction of Bisdioxopiperazines with Topoisomerase II{alpha} in Vivo
Mol. Pharmacol., October 1, 2007; 72(4): 1003 - 1014.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
L. H. Jensen, A. V. Thougaard, M. Grauslund, B. Sokilde, E. V. Carstensen, H. K. Dvinge, D. A. Scudiero, P. B. Jensen, R. H. Shoemaker, and M. Sehested
Substituted Purine Analogues Define a Novel Structural Class of Catalytic Topoisomerase II Inhibitors
Cancer Res., August 15, 2005; 65(16): 7470 - 7477.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Vaughn, S. Huang, I. Wessel, T. K. Sorensen, T. Hsieh, L. H. Jensen, P. B. Jensen, M. Sehested, and J. L. Nitiss
Stability of the Topoisomerase II Closed Clamp Conformation May Influence DNA-stimulated ATP Hydrolysis
J. Biol. Chem., March 25, 2005; 280(12): 11920 - 11929.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. V. Walker, K. C. Nitiss, L. H. Jensen, C. Mayne, T. Hu, P. B. Jensen, M. Sehested, T. Hsieh, and J. L. Nitiss
A Mutation in Human Topoisomerase II {alpha} Whose Expression Is Lethal in DNA Repair-deficient Yeast Cells
J. Biol. Chem., June 18, 2004; 279(25): 25947 - 25954.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
N. Vilain, M. Tsai-Pflugfelder, A. Benoit, S. M. Gasser, and D. Leroy
Modulation of drug sensitivity in yeast cells by the ATP-binding domain of human DNA topoisomerase II{alpha}
Nucleic Acids Res., October 1, 2003; 31(19): 5714 - 5722.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. Classen, S. Olland, and J. M. Berger
Structure of the topoisomerase II ATPase region and its mechanism of inhibition by the chemotherapeutic agent ICRF-187
PNAS, September 16, 2003; 100(19): 10629 - 10634.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
A. Renodon-Corniere, T. K. Sorensen, P. B. Jensen, J. L. Nitiss, B. Sokilde, M. Sehested, and L. H. Jensen
Probing the Role of Linker Substituents in Bisdioxopiperazine Analogs for Activity against Wild-Type and Mutant Human Topoisomerase IIalpha
Mol. Pharmacol., May 1, 2003; 63(5): 1159 - 1168.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. Skouboe, L. Bjergbaek, V. H. Oestergaard, M. K. Larsen, B. R. Knudsen, and A. H. Andersen
A Human Topoisomerase IIalpha Heterodimer with Only One ATP Binding Site Can Go through Successive Catalytic Cycles
J. Biol. Chem., February 14, 2003; 278(8): 5768 - 5774.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
L. H. Jensen, A. Renodon-Corniere, I. Wessel, S. W. Langer, B. Sokilde, E. V. Carstensen, M. Sehested, and P. B. Jensen
Maleimide Is a Potent Inhibitor of Topoisomerase II in Vitro and in Vivo: A New Mode of Catalytic Inhibition
Mol. Pharmacol., May 1, 2002; 61(5): 1235 - 1243.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. Hu, H. Sage, and T.-s. Hsieh
ATPase Domain of Eukaryotic DNA Topoisomerase II. INHIBITION OF ATPase ACTIVITY BY THE ANTI-CANCER DRUG BISDIOXOPIPERAZINE AND ATP/ADP-INDUCED DIMERIZATION
J. Biol. Chem., February 15, 2002; 277(8): 5944 - 5951.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
B. B. Hasinoff, M. E. Abram, N. Barnabé, T. Khélifa, W. P. Allan, and J. C. Yalowich
The Catalytic DNA Topoisomerase II Inhibitor Dexrazoxane (ICRF-187) Induces Differentiation and Apoptosis in Human Leukemia K562 Cells
Mol. Pharmacol., March 1, 2001; 59(3): 453 - 461.
[Abstract] [Full Text]

Home page
 J. Pharmacol. Exp. Ther.Home page
B. B. Hasinoff, M. E. Abram, G.-L. Chee, E. Huebner, E. H. Byard, N. Barnabé, V. J. Ferrans, Z.-X. Yu, and J. C. Yalowich
The Catalytic DNA Topoisomerase II Inhibitor Dexrazoxane (ICRF-187) Induces Endopolyploidy in Chinese Hamster Ovary Cells
J. Pharmacol. Exp. Ther., November 1, 2000; 295(2): 474 - 483.
[Abstract] [Full Text]

Home page
 Mol. Pharmacol.Home page
S. Patel, E. Jazrawi, A. M. Creighton, C. A. Austin, and L. M. Fisher
Probing the Interaction of the Cytotoxic Bisdioxopiperazine ICRF-193 with the Closed Enzyme Clamp of Human Topoisomerase IIalpha
Mol. Pharmacol., September 1, 2000; 58(3): 560 - 568.
[Abstract] [Full Text]

Home page
 Clin. Cancer Res.Home page
S. W. Langer, M. Sehested, and P. B. Jensen
Treatment of Anthracycline Extravasation with Dexrazoxane
Clin. Cancer Res., September 1, 2000; 6(9): 3680 - 3686.
[Abstract] [Full Text]

Home page
 Mol. Pharmacol.Home page
S. E. Morgan, R. S. Cadena, S. C. Raimondi, and W. T. Beck
Selection of Human Leukemic CEM Cells for Resistance to the DNA Topoisomerase II Catalytic Inhibitor ICRF-187 Results in Increased Levels of Topoisomerase IIalpha and Altered G2/M Checkpoint and Apoptotic Responses
Mol. Pharmacol., February 1, 2000; 57(2): 296 - 307.
[Abstract] [Full Text]

Home page
 J. Biol. Chem.Home page
S. K. Morris, C. L. Baird, and J. E. Lindsley
Steady-state and Rapid Kinetic Analysis of Topoisomerase II Trapped as the Closed-clamp Intermediate by ICRF-193
J. Biol. Chem., January 28, 2000; 275(4): 2613 - 2618.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. Tsutsui, K. Tsutsui, K. Sano, A. Kikuchi, and A. Tokunaga
Involvement of DNA Topoisomerase IIbeta in Neuronal Differentiation
J. Biol. Chem., February 16, 2001; 276(8): 5769 - 5778.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online
Copyright © 1999 by the American Association for Cancer Research.