Molecular Analysis of Programmed Cell Death by DNA Topoisomerase Inhibitors in Kinetoplastid Parasite Leishmania

  • Nilkantha Sen
  • Bijoylaxmi Banerjee
  • Hemanta K. Majumder
Part of the Molecular Biology Intelligence Unit book series (MBIU)


DNA topoisomerases of kinetoplastid parasites represent a family of DNA processing enzymes that essentially solve the topological problems not only in nuclear DNA but also in kinetoplastid DNA. Due to their indispensable function in cell biology they are valuable potential targets for many antileishmanial drugs to induce programmed cell death (PCD). PCD is thought to have evolved to regulate growth and development of multicellular organisms. Kinetoplastid organisms have developed an altruistic mechanism to promote and maintain the clonality within their population. Characterization of PCD by topoisomerase inhibitors could provide information regarding their pathogenesis which could be exploited to develop new targets to limit their growth and treat the disease they cause.

DNA topology affects fundamental processes of life and DNA topoisomerases constitute a growing family of nuclear enzymes that regulate unconstrained DNA supercoils in all living species, bacteria, archea, kinetoplastids and eukaryotes. Changes in DNA topology are required for virtually all DNA dependent events such as DNA replication, transcription, recombination, repair, nucleosome remodeling, chromosome condensation and segregation.1,2 Topoisomerases are divided into two classes, based primarily on their mode of cleaving DNA. Type I DNA topoisomerases act by making a transient nick on a single strand of duplex DNA, passing another strand through the nick and changing the linking number by one unit.3 Type II topoisomerases act by transiently nicking both strands of the DNA, passing another double stranded DNA segment through the gap and changing the linking number by two with the help of ATP molecules.4


Trypanosoma Brucei Death Induce Signal Complex Cleavage Complex Leishmania Donovani Kinetoplastid Parasite 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Wang JC. Cellular roles of DNA topoisomerases: a molecular perspective. Nat Rev Mol Cell Biol 2002; 3:430–440.PubMedCrossRefGoogle Scholar
  2. 2.
    Cortes F, Pastor N, Mateos S et al. Roles of DNA topoisomerases in chromosome segregation and mitosis. Mutat Res 2003; 543:59–66.PubMedCrossRefGoogle Scholar
  3. 3.
    Stewart L, Ireton GC, Champoux JJ. Reconstitution of human topoisomerase I by fragment complementation. J Mol Biol 1997; 269:355–372.PubMedCrossRefGoogle Scholar
  4. 4.
    Berger JM. Structure of DNA topoisomerases. Biochim Biophys Acta 1998; 1400:3–18.PubMedGoogle Scholar
  5. 5.
    Das BB, Sen N, Ganguly A et al. Reconstitution and functional characterization of the unusual bi-subunit type I DNA topoisomerase from Leishmania donovani. FEBS Lett 2004; 565:81–88.PubMedCrossRefGoogle Scholar
  6. 6.
    Villa H, Otero Marcos AR, Reguera RM et al. A novel active DNA topoisomerase I in Leishmania donovani. J Biol Chem 2003; 278:3521–3526.PubMedCrossRefGoogle Scholar
  7. 7.
    Das BB, Sen N, Dasgupta SB et al. N-terminal region of the large subunit of Leishmania donovani bisubunit topoisomerase I is involved in DNA relaxation and interaction with the smaller subunit. J Biol Chem 2005; 280;16335–16344.PubMedCrossRefGoogle Scholar
  8. 8.
    Das BB, Sen N, Roy A et al. Differential induction of Leishmania donovani bi-subunit topoisomerase I-DNA cleavage complex by selected flavones and camptothecin: activity of flavones against camptothecin-resistant topoisomerase I. Nucleic Acids Res 2006; 34:1121–1132.PubMedCrossRefGoogle Scholar
  9. 9.
    Cheng C, Kussie P, Pavletich N et al. Conservation of structure and mechanism between eukaryotic topoisomerase I and site-specific recombinases. Cell 1998; 92:841–850.PubMedCrossRefGoogle Scholar
  10. 10.
    Pasion SG, Hines JC, Aebersold R et al. Molecular cloning and expression of the gene encoding the kinetoplast-associated type II DNA topoisomerase of Crithidia fasciculata. Mol Biochem Parasitol 1992; 50:57–67.PubMedCrossRefGoogle Scholar
  11. 11.
    Das A, Dasgupta A, Sharma S et al. Characterisation of the gene encoding type II DNA topoisomerase from Leishmania donovani: a key molecular target in antileishmanial therapy. Nucleic Acids Res 2001; 29:1844–1851.PubMedCrossRefGoogle Scholar
  12. 12.
    Strauss PR, Wang JC. The TOP2 gene of Trypanosoma brucei: a single-copy gene that shares extensive homology with other TOP2 genes encoding eukaryotic DNA topoisomerase II. Mol Biochem Parasitol 1990; 38:141–150.PubMedCrossRefGoogle Scholar
  13. 13.
    Fragoso SP, Goldenberg S. Cloning and characterization of the gene encoding Trypanosoma cruzi DNA topoisomerase II. Mol Biochem Parasitol 1992; 55:127–134.PubMedCrossRefGoogle Scholar
  14. 14.
    Gaziova I, Lukes J. Mitochondrial and nuclear localization of topoisomerase II in the flagellate Bodo saltans (Kinetoplastida), a species with noncatenated kinetoplast DNA. J Biol Chem 2003; 278:10900–10907.PubMedCrossRefGoogle Scholar
  15. 15.
    Sengupta T, Mukherjee M, Mandal C et al. Functional dissection of the C-terminal domain of type II DNA topoisomerase from the kinetoplastid hemoflagellate Leishmania donovani. Nucleic Acids Res 2003; 31:5305–5316.PubMedCrossRefGoogle Scholar
  16. 16.
    Das A, Mandal C, Dasgupta A et al. An insight into the active site of a type I DNA topoisomerase from the kinetoplastid protozoan Leishmania donovani. Nucleic Acids Res 2002; 30:794–802.PubMedCrossRefGoogle Scholar
  17. 17.
    Sengupta T, Mukherjee M, Das R et al. Characterization of the DNA-binding domain and identification of the active site residue in the ‘Gyr A’ half of Leishmania donovani topoisomerase II. Nucleic Acids Res 2005; 33:2364–2373.PubMedCrossRefGoogle Scholar
  18. 18.
    Ferguson M, Torri AF, Perez-Morga Ward DC et al. DNA replication: mechanistic differences between Trypanosoma brucei and Crithidia fasciculata. J Cell Biol 1994; 126:631–639.PubMedCrossRefGoogle Scholar
  19. 19.
    Wang Z, Drew ME, Morris JC et al. Asymmetrical division of the kinetoplast DNA network of the trypanosome. EMBO J 2002; 21:4998–5005.PubMedCrossRefGoogle Scholar
  20. 20.
    Myler PJ, Glick D, Feagin JE et al. Structural organization of the maxicircle variable region of Trypanosoma brucei: identification of potential replication origins and topoisomerase II binding sites. Nucleic Acids Res 1993; 21:687–694.PubMedCrossRefGoogle Scholar
  21. 21.
    Sloof P, deHaan A, Eier W et al. The nucleotide sequence of the variable region in Trypanosoma brucei completes the sequence analysis of the maxicircle component of mitochondrial kinetoplast DNA. Mol Biochem Parasitol 1992; 56:289–299.PubMedCrossRefGoogle Scholar
  22. 22.
    Hajduk Sl, Klein VA, Englund PT. Replication of kinetoplast DNA maxicircles Cell 1984; 36:483–492.PubMedCrossRefGoogle Scholar
  23. 23.
    Perez-Morga D, Englund PT. The structure of replicating kinetoplast DNA networks. J Cell Biol 1993; 123:1069–1079.PubMedCrossRefGoogle Scholar
  24. 24.
    Wang JC. DNA topoisomerases as targets of therapeutics: an overview. Adv Pharmacol 1994; 29A:1–19.PubMedCrossRefGoogle Scholar
  25. 25.
    Ulukan H, Swaan PW. Camptothecins: a review of their chemotherapeutic potential. Drugs 2002; 62:2039–2057.PubMedCrossRefGoogle Scholar
  26. 26.
    Kim DK, Lee N. Recent advances in topoisomerase I-targeting agents, camptothecin analogues. Mini Rev Med Chem 2002; 2:611–619.PubMedCrossRefGoogle Scholar
  27. 27.
    Grabowski DR, Holmes KA, Aoyama M et al. Altered drug interaction and regulation of topoisomerase IIbeta: potential mechanisms governing sensitivity of HL-60 cells to amsacrine and etoposide. Mol Pharmacol 1999; 56:1340–1345.PubMedGoogle Scholar
  28. 28.
    Snyder RD. Evidence from studies with intact mammalian cells that merbarone and bis(dioxopiperazine)s are topoisomerase II poisons. Drug Chem Toxicol 2003; 26:15–22.PubMedCrossRefGoogle Scholar
  29. 29.
    Lewy DS, Gauss CM, Soenen DR et al. Fostriecin: chemistry and biology. Curr Med Chem 2002; 9:2005–2032.PubMedGoogle Scholar
  30. 30.
    Sordet O, Khan QA, Kohn KW et al. Apoptosis induced by topoisomerase inhibitors. Curr Med Chem Anticancer Agents 2003; 3:271–290.PubMedCrossRefGoogle Scholar
  31. 31.
    Mills JC, Stone NL, Pittman RN. Extranuclear apoptosis. The role of the cytoplasm in the execution phase. J Cell Biol 1999; 146:703–708.PubMedCrossRefGoogle Scholar
  32. 32.
    Wanderley JL, Benjamin A, Real F et al. Apoptotic mimicry: an altruistic behavior in host/Leishmania interplay. Braz J Med Biol Res 2005; 38:807–812.PubMedCrossRefGoogle Scholar
  33. 33.
    Debrabant A, Lee N, Bertholet S et al. Programmed cell death in trypanosomatids and other unicellular organisms. Int J Parasitol 2003; 33:257–267.PubMedCrossRefGoogle Scholar
  34. 34.
    Szallies A, Kubata BK, Duszenko M. A metacaspase of Trypanosoma brucei causes loss of respiration competence and clonal death in the yeast Saccharomyces cerevisiae. FEBS Lett 2002; 517:144–150.PubMedCrossRefGoogle Scholar
  35. 35.
    Luo J, Nikolaev AY, Imai S et al. Negative control of p53 by Sir2alpha promotes cell survival under stress. Cell 2001; 107:137–148.PubMedCrossRefGoogle Scholar
  36. 36.
    Duttaroy A, Bourbeau D, Wang XL et al. Apoptosis rate can be accelerated or decelerated by overexpression or reduction of the level of elongation factor-1 alpha. Exp Cell Res 1998; 238:168–176.PubMedCrossRefGoogle Scholar
  37. 37.
    Solari E, Droin N, Batteiab A et al. Positive and negative regulation of apoptotic pathways by cytotoxic agents in hematological malignancies. Leukemia 2000; 14:1833–1849.CrossRefGoogle Scholar
  38. 38.
    Ravagnan L, Roumier T, Kroemer G. Mitochondria, the killer organelles and their weapons. J Cell Physiol 2002; 192:131–137.PubMedCrossRefGoogle Scholar
  39. 39.
    Nagata S. Apoptosis by death factor. Cell 1997; 88:355–365.PubMedCrossRefGoogle Scholar
  40. 40.
    Salvesen GS, Dixit VM. Caspase activation: the induced-proximity model. Proc Natl Acad Sci 1999; 96:10964–10967.PubMedCrossRefGoogle Scholar
  41. 41.
    Sen N, Das BB, Ganguly A et al. Camptothecin induced mitochondrial dysfunction leading to programmed cell death in unicellular hemoflagellate Leishmania donovani. Cell Death Differ 2004; 11:924–936.PubMedCrossRefGoogle Scholar
  42. 42.
    Sen N, Das BB, Ganguly A et al. Camptothecin-induced imbalance in intracellular cation homeostasis regulates programmed cell death in unicellular hemoflagellate Leishmania donovani. J Biol Chem 2004; 279:52366–52375.PubMedCrossRefGoogle Scholar
  43. 43.
    Mittra B, Saha A, Chowdhury AR et al. Luteolin, an abundant dietary component is a potent anti-leishmanial agent that acts by inducing topoisomerase II-mediated kinetoplast DNA cleavage leading to apoptosis. Mol Med 2000; 6:527–541.PubMedCrossRefGoogle Scholar
  44. 44.
    Sen N, Das BB, Ganguly A et al. Intracellular ATP level regulates apoptosis-like death in luteolin induced dyskinetoplastid cells. Exp Parasitol 2006.Google Scholar
  45. 45.
    Singh G, Jayanarayan KG, Dey CS. Novobiocin induces apoptosis-like cell death in topoisomerase II over-expressing arsenite resistant Leishmania donovani. Mol Biochem Parasitol 2005; 141:57–69.PubMedCrossRefGoogle Scholar
  46. 46.
    Ouaissi A. Apoptosis-like death in trypanosomatids: search for putative pathways and genes involved. Kinetoplastid Biol Dis, 2003:2–5.Google Scholar
  47. 47.
    Yang SW, Burgin AB Jr, Huizenga BN et al. A eukaryotic enzyme that can disjoin dead-end covalent complexes between DNA and type I topoisomerases. Proc Natl Acad Sci 1996; 93:11534–11539.PubMedCrossRefGoogle Scholar

Copyright information

© Landes Bioscience and Springer Science+Business Media 2008

Authors and Affiliations

  • Nilkantha Sen
    • 1
  • Bijoylaxmi Banerjee
    • 1
  • Hemanta K. Majumder
    • 1
  1. 1.Division of Molecular ParasitologyIndian Institute of Chemical BiologyKolkataIndia

Personalised recommendations