ACS Publications. Most Trusted. Most Cited. Most Read
My Activity
Recently Viewed
You have not visited any articles yet, Please visit some articles to see contents here.

Figure 1Loading Img

Mutations of Human Topoisomerase IIα Affecting Multidrug Resistance and Sensitivity

View Author Information
Department of Pharmacology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, New Jersey 08854, Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710, and Department of Molecular Pharmacology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101
Cite this: Biochemistry 1999, 38, 33, 10793–10800
Publication Date (Web):July 29, 1999
Copyright © 1999 American Chemical Society
Article Views
Read OnlinePDF (146 KB)


Two mutations, R450Q and P803S, in the coding region of the human topoisomerase IIα gene have been identified in the atypical multidrug resistant (at-MDR) cell line, CEM/VM-1, which exhibits resistance to many structurally diverse topoisomerase II-targeting antitumor drugs such as VM-26, doxorubicin, m-AMSA, and mitoxantrone. The R450Q mutation mapped in the ATP utilization domain, while the P803S mutation mapped in the vicinity of the active site tyrosine of human topoisomerase IIα. However, the roles of these two mutations in conferring multidrug resistance are unclear. To study the roles of these two mutations in conferring multidrug resistance, we have characterized the recombinant human DNA topoisomerase IIα containing either single or double mutations. We show that both R450Q and P803S mutations confer resistance in the absence of ATP. However, in the presence of ATP, the R450Q, but not the P803S, mutation can confer multidrug resistance. The R450Q enzyme was shown to exhibit impaired ATP utilization both for enzyme catalysis and for its ability to form the circular protein clamp. Interestingly, an unrelated mutation, G437E, which is also located in the same domain as the R450Q mutation, exhibited multidrug hypersensitivity in the absence of ATP. However, in the presence of ATP, the G437E enzyme is only minimally hypersensitive to various topoisomerase II drugs. In contrast to the R450Q enzyme, the G437E enzyme exhibited enhanced ATP utilization for enzyme catalysis. In the aggregate, these results support the notion that the multidrug resistance and sensitivity of these mutant enzymes are due to a specific defect in ATP utilization during enzyme catalysis.

 This work was supported by NIH Grants CA39962 and GM27731 (to L.F.L.), CA21765 and CA52814 (to J.L.N.), and GM29006 (to T.-S.H.).

 University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School.


 Duke University Medical Center.

 St. Jude Children's Research Hospital.


 To whom correspondence should be addressed:  Department of Pharmacology, UMDNJ-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, NJ 08854. Telephone:  (732) 235-4592. Fax:  (732) 235-4073. E-mail:  [email protected]

Cited By

This article is cited by 37 publications.

  1. Yunke Wang, Samantha Rakela, Jeremy W. Chambers, Zi-Chun Hua, Mark T. Muller, John L. Nitiss, Yuk-Ching Tse-Dinh, Fenfei Leng. Kinetic Study of DNA Topoisomerases by Supercoiling-Dependent Fluorescence Quenching. ACS Omega 2019, 4 (19) , 18413-18422.
  2. May Levin, Michal Stark, Yishai Ofran, Yehuda G. Assaraf. Deciphering molecular mechanisms underlying chemoresistance in relapsed AML patients: towards precision medicine overcoming drug resistance. Cancer Cell International 2021, 21 (1)
  3. Sandhya Bansal, Priyanka Bajaj, Stuti Pandey, Vibha Tandon. Topoisomerases: Resistance versus Sensitivity, How Far We Can Go?. Medicinal Research Reviews 2017, 37 (2) , 404-438.
  4. Prashanth Gokare, Arunasalam Navaraj, Shengliang Zhang, Noboru Motoyama, Shen-Shu Sung, Niklas K. Finnberg. Targeting of Chk2 as a countermeasure to dose-limiting toxicity triggered by topoisomerase-II (TOP2) poisons. Oncotarget 2016, 7 (20) , 29520-29530.
  5. Hiroyuki Tamaichi, Masaki Sato, Andrew C. G. Porter, Toshiaki Shimizu, Shuki Mizutani, Masatoshi Takagi. Ataxia telangiectasia mutated-dependent regulation of topoisomerase II alpha expression and sensitivity to topoisomerase II inhibitor. Cancer Science 2013, 104 (2) , 178-184.
  6. Manal M. Zyada, Nagla M. Salama. Detection of the Epstein-Barr Virus and DNA-Topoisomerase II- α in Recurrent and Nonrecurrent Giant Cell Lesion of the Jawbones. BioMed Research International 2013, 2013 , 1-5.
  7. Anand G Patel, Scott H Kaufmann. How does doxorubicin work?. eLife 2012, 1
  8. Anna Rogojina, Stefan Gajewski, Karim Bahmed, Neil Osheroff, John L. Nitiss. Topoisomerase II Inhibitors: Chemical Biology. 2012,,, 211-243.
  9. David Jamieson, Alan V Boddy. Pharmacogenetics of genes across the doxorubicin pathway. Expert Opinion on Drug Metabolism & Toxicology 2011, 7 (10) , 1201-1210.
  10. Stephen J. Cutler, Horace G. Cutler. Natural Products as Cytotoxic Agents. 2010,,, 1-48.
  11. Jonathan Sperry, Isabel Lorenzo-Castrillejo, Margaret A. Brimble, Felix Machín. Pyranonaphthoquinone derivatives of eleutherin, ventiloquinone L, thysanone and nanaomycin A possessing a diverse topoisomerase II inhibition and cytotoxicity spectrum. Bioorganic & Medicinal Chemistry 2009, 17 (20) , 7131-7137.
  12. Andrew C.G. Porter. Depletion and Mutation of Topoisomerase II in Animal Cells. 2009,,, 245-263.
  13. Gaganmeet Singh, Meghna Thakur, Pradip K. Chakraborti, Chinmoy S. Dey. Evidence for the presence of R250G mutation at the ATPase domain of topoisomerase II in an arsenite-resistant Leishmania donovani exhibiting a differential drug inhibition profile. International Journal of Antimicrobial Agents 2009, 33 (1) , 80-85.
  14. Anna T. Rogojina, John L. Nitiss. Isolation and Characterization of mAMSA-hypersensitive Mutants. Journal of Biological Chemistry 2008, 283 (43) , 29239-29250.
  15. Darren J. Burgess, Jason Doles, Lars Zender, Wen Xue, Beicong Ma, W. Richard McCombie, Gregory J. Hannon, Scott W. Lowe, Michael T. Hemann. Topoisomerase levels determine chemotherapy response in vitro and in vivo. Proceedings of the National Academy of Sciences 2008, 105 (26) , 9053-9058.
  16. Gaetano Marverti, Matteo Cusumano, Alessio Ligabue, Maria Letizia Di Pietro, Pasquale Antonio Vainiglia, Angela Ferrari, Margherita Bergomi, Maria Stella Moruzzi, Chiara Frassineti. Studies on the anti-proliferative effects of novel DNA-intercalating bipyridyl–thiourea–Pt(II) complexes against cisplatin-sensitive and -resistant human ovarian cancer cells. Journal of Inorganic Biochemistry 2008, 102 (4) , 699-712.
  17. Haiyan Qi, Chao-Po Lin, Xuan Fu, Laurence M. Wood, Angela A. Liu, Yuan-Chin Tsai, Yongjie Chen, Christopher M. Barbieri, Daniel S. Pilch, Leroy F. Liu. G-Quadruplexes Induce Apoptosis in Tumor Cells. Cancer Research 2006, 66 (24) , 11808-11816.
  18. Linda Rickardson, Mårten Fryknäs, Caroline Haglund, Henrik Lövborg, Peter Nygren, Mats G. Gustafsson, Anders Isaksson, Rolf Larsson. Screening of an annotated compound library for drug activity in a resistant myeloma cell line. Cancer Chemotherapy and Pharmacology 2006, 58 (6) , 749-758.
  19. Mårten Fryknäs, Linda Rickardson, Malin Wickström, Sumeer Dhar, Henrik Lövborg, Joachim Gullbo, Peter Nygren, Mats G. Gustafsson, Anders Isaksson, Rolf Larsson. Phenotype-Based Screening of Mechanistically Annotated Compounds in Combination with Gene Expression and Pathway Analysis Identifies Candidate Drug Targets in a Human Squamous Carcinoma Cell Model. SLAS Discovery 2006, 11 (5) , 457-468.
  20. L Rickardson, M Fryknäs, S Dhar, H Lövborg, J Gullbo, M Rydåker, P Nygren, M G Gustafsson, R Larsson, A Isaksson. Identification of molecular mechanisms for cellular drug resistance by combining drug activity and gene expression profiles. British Journal of Cancer 2005, 93 (4) , 483-492.
  21. Xiaofeng Jiang. Random mutagenesis of the B′A′ core domain of yeast DNA topoisomerase II and large-scale screens of mutants resistant to the anticancer drug etoposide. Biochemical and Biophysical Research Communications 2005, 327 (2) , 597-603.
  22. Jerrylaine V. Walker, Karin C. Nitiss, Lars H. Jensen, Christopher Mayne, Tao Hu, Peter B. Jensen, Maxwell Sehested, Tao Hsieh, John L. Nitiss. A Mutation in Human Topoisomerase II α Whose Expression Is Lethal in DNA Repair-deficient Yeast Cells. Journal of Biological Chemistry 2004, 279 (25) , 25947-25954.
  23. Vibe H. Oestergaard, Lotte Bjergbaek, Camilla Skouboe, Laura Giangiacomo, Birgitta R. Knudsen, Anni H. Andersen. The Transducer Domain Is Important for Clamp Operation in Human DNA Topoisomerase IIα. Journal of Biological Chemistry 2004, 279 (3) , 1684-1691.
  24. Lester A. Mitscher, Apurba Dutta. Antitumor Natural Products. 2003,,, 107-150.
  25. Leroy F. Liu. Degradation of topoisomerase cleavable complexes. 2003,,, 79-88.
  26. Mobeen Malik, John L. Nitiss. Understanding the action of drugs targeting TOP2: Yeast based systems for unraveling drug mechanisms. 2003,,, 109-127.
  27. Heidi Schwarzenbach. A diagnostic tool for monitoring multidrug resistance expression in human tumor tissues. Analytical Biochemistry 2002, 308 (1) , 26-33.
  28. Irene Wessel, Lars H. Jensen, Axelle Renodon-Corniere, Tina K. Sorensen, John L. Nitiss, Peter B. Jensen, Maxwell Sehested. Human small cell lung cancer NYH cells resistant to the bisdioxopiperazine ICRF-187 exhibit a functional dominant Tyr165Ser mutation in the Walker A ATP binding site of topoisomerase IIα. FEBS Letters 2002, 520 (1-3) , 161-166.
  29. Z. Topcu. DNA topoisomerases as targets for anticancer drugs. Journal of Clinical Pharmacy and Therapeutics 2001, 26 (6) , 405-416.
  30. Yoshito Okada, Aki Tosaka, Yuji Nimura, Akihiko Kikuchi, Shonen Yoshida, Motoshi Suzuki. Atypical multidrug resistance may be associated with catalytically active mutants of human DNA topoisomerase II α. Gene 2001, 272 (1-2) , 141-148.
  31. John M. Fortune, Oleg V. Lavrukhin, James R. Gurnon, James L. Van Etten, R. Stephen Lloyd, Neil Osheroff. Topoisomerase II from Chlorella Virus PBCV-1 Has an Exceptionally High DNA Cleavage Activity. Journal of Biological Chemistry 2001, 276 (26) , 24401-24408.
  32. Huimin Wang, Yong Mao, Nai Zhou, Tao Hu, Tao-Shih Hsieh, Leroy F. Liu. ATP-bound Topoisomerase II as a Target for Antitumor Drugs. Journal of Biological Chemistry 2001, 276 (19) , 15990-15995.
  33. Tsai-Kun Li, Leroy F Liu. Tumor Cell Death Induced by Topoisomerase-Targeting Drugs. Annual Review of Pharmacology and Toxicology 2001, 41 (1) , 53-77.
  34. Alexander Arlt, Jens Vorndamm, Maike Breitenbroich, Ulrich R Fölsch, Holger Kalthoff, Wolfgang E Schmidt, Heiner Schäfer. Inhibition of NF-κB sensitizes human pancreatic carcinoma cells to apoptosis induced by etoposide (VP16) or doxorubicin. Oncogene 2001, 20 (7) , 859-868.
  35. Yong Mao, Shyamal D. Desai, Leroy F. Liu. SUMO-1 Conjugation to Human DNA Topoisomerase II Isozymes. Journal of Biological Chemistry 2000, 275 (34) , 26066-26073.
  36. Yoshito Okada, Yasutomo Ito, Akihiko Kikuchi, Yuji Nimura, Shonen Yoshida, Motoshi Suzuki. Assignment of Functional Amino Acids around the Active Site of Human DNA Topoisomerase IIα. Journal of Biological Chemistry 2000, 275 (32) , 24630-24638.
  37. Giuseppe Giaccone, Giovanni Capranico. 10th Conference on DNA Topoisomerases in therapy. Drug Resistance Updates 1999, 2 (6) , 347-350.

Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

You’ve supercharged your research process with ACS and Mendeley!

Click to create an ACS ID

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Your Mendeley pairing has expired. Please reconnect

This website uses cookies to improve your user experience. By continuing to use the site, you are accepting our use of cookies. Read the ACS privacy policy.