Thymoquinone disrupts the microtubule dynamics in fission yeast <em>Schizosaccharomyces pombe</em>

Nusrat Masood; Saman Khan; Suaib Luqman; Shakil Ahmed; Nusrat Masood; Saman Khan; Suaib Luqman; Shakil Ahmed

doi:10.3934/genet.2016.4.239

AIMS Genetics

2016, Volume 3, Issue 4: 239-251. doi: 10.3934/genet.2016.4.239

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Research article

Thymoquinone disrupts the microtubule dynamics in fission yeast Schizosaccharomyces pombe

1.
Molecular Bioprospection Department of Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow-226015, Uttar Pradesh, India.
2.
Molecular and Structural Biology Division, CSIR—Central Drug Research Institute, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow-226031, India,

Received: 29 August 2016 Accepted: 11 November 2016 Published: 16 November 2016

Mad2 deletion strain of Schizosaccharomyces pombe was found to be sensitive to thymoquinone, a signature molecule present in Nigella sativa in a dose-dependent manner. Mad2 protein is an indispensable part of mitotic spindle checkpoint complex and is required for the cell cycle arrest in response to the spindle defects. Although the expression of α tubulin was not affected in thymoquinone treated cells, but the expression of β-tubulin was reduced. Further, the absence of microtubule in thymoquinone treated cells suggests its involvement in tubulin polymerization. Molecular docking studies revealed that thymoquinone specifically binds to β-tubulin near the Taxotere binding site of Tub1 (Tubulin α-β dimer). These studies additionally showed that thymoquinone interacts with the residues present in chain B, which is an inherent part of Mad2 protein of mitotic checkpoint complex (MCC). We concluded that the thymoquinone disrupts the microtubule polymerization that leads to the requirement of spindle checkpoint protein for the cell survival.
- Thymoquinone,
- spindle assembly checkpoint,
- Mad2,
- Tubulin
Citation: Nusrat Masood, Saman Khan, Suaib Luqman, Shakil Ahmed. Thymoquinone disrupts the microtubule dynamics in fission yeast Schizosaccharomyces pombe[J]. AIMS Genetics, 2016, 3(4): 239-251. doi: 10.3934/genet.2016.4.239

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Abstract

Mad2 deletion strain of Schizosaccharomyces pombe was found to be sensitive to thymoquinone, a signature molecule present in Nigella sativa in a dose-dependent manner. Mad2 protein is an indispensable part of mitotic spindle checkpoint complex and is required for the cell cycle arrest in response to the spindle defects. Although the expression of α tubulin was not affected in thymoquinone treated cells, but the expression of β-tubulin was reduced. Further, the absence of microtubule in thymoquinone treated cells suggests its involvement in tubulin polymerization. Molecular docking studies revealed that thymoquinone specifically binds to β-tubulin near the Taxotere binding site of Tub1 (Tubulin α-β dimer). These studies additionally showed that thymoquinone interacts with the residues present in chain B, which is an inherent part of Mad2 protein of mitotic checkpoint complex (MCC). We concluded that the thymoquinone disrupts the microtubule polymerization that leads to the requirement of spindle checkpoint protein for the cell survival.

References

[1]	Simon JA, Bedalov A (2004) Yeast as a model system for anticancer drug discovery. Nat Rev Cancer 4: 481-487.
[2]	Habu T, Matsumoto T (2013) p31 (comet) inactivates the chemically induced Mad2-dependent spindle assembly checkpoint and leads to resistance to anti-mitotic drugs. Springer Plus 2: 562. doi: 10.1186/2193-1801-2-562
[3]	Elledge SJ (1998) Mitotic arrest: Mad2 prevents sleepy from waking up the APC. Science 279: 999-1000.
[4]	Kops GL, Weaver BA, Cleveland DW (2005) On the road to cancer: aneuploidy and the mitotic checkpoint. Nat Rev Cancer 5: 773-785.
[5]	Nasmyth K (2005) How do so few control so many? Cell 120: 739-746.
[6]	Page BD, Snyder M (1993) Chromosome segregation in yeast. Annu Rev Microbiol 47: 231-261. doi: 10.1146/annurev.mi.47.100193.001311
[7]	Amon A (1999) The spindle checkpoint. Curr Opin Genet Dev 9: 69-75.
[8]	Lew DJ, Burke DJ (2003) The Spindle Assembly and Spindle Position Checkpoints. Annu Rev Genet 37: 251-282.
[9]	Pinsky BA, Biggins S (2005) The spindle checkpoint: tension versus attachment. Trends Cell Biol 15: 486-493. doi: 10.1016/j.tcb.2005.07.005
[10]	Li R, Murray AW (1991) Feedback control of mitosis in budding yeast. Cell 66: 519-531.
[11]	Hoyt MA, Totis L, Roberts BT (1991) S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function. Cell 66: 507-517.
[12]	Foley EA, Kapoor TM (2013) Microtubule attachment and spindle assembly checkpoint signalling at the kinetochore. Nat Rev Mol Cell Biol 14: 25-37.
[13]	Schweizer N, Ferras C, Kern DM, et al. (2013) Spindle assembly checkpoint robustness requires Tpr-mediated regulation of Mad1/Mad2 proteostasis. J Cell Biol 203: 883-893.
[14]	Musacchio A, Salmon ED (2007) The spindle-assembly checkpoint in space and time. Nat Rev Mol Cell Biol 8: 379-393.
[15]	He X, Jones MH, Winey M, et al. (1998) Mph1, a member of the Mps1-like family of dual specificity protein kinases, is required for the spindle checkpoint in S. pombe. J Cell Sci 111: 1635-1647.
[16]	He X, Patterson TE, Sazer S (1997) The Schizosaccharomyces pombe spindle checkpoint protein mad2p blocks anaphase and genetically interacts with the anaphase-promoting complex. Proc Natl Acad Sci USA 94: 7965-7970.
[17]	Rajagopalan H, Lengauer C (2004) Aneuploidy and cancer. Nature 432: 338-341.
[18]	Pandit SK, Westendorp B, de Bruin A (2013) Physiological significance of polyploidization in mammalian cells. Trends Cell Biol 23: 556-566.
[19]	Severina II, Severin FF, Korshunova GA, et al. (2013) In search of novel highly active mitochondria-targeted antioxidants: thymoquinone and its cationic derivatives. FEBS Lett 587: 2018-2024.
[20]	Yu SM, Kim SJ (2013) Thymoquinone-induced reactive oxygen species causes apoptosis of chondrocytes via PI3K/Akt and p38kinase pathway. Exp Biol Med 238: 811-820. doi: 10.1177/1535370213492685
[21]	Alhosin IM, Ibrahim A, Boukhari A, et al. (2012) Anti-neoplastic agent thymoquinone induces degradation of alpha and beta tubulin proteins in human cancer cells without affecting their level in normal human fibroblasts. Invest New Drugs 30: 1813-1819.
[22]	Verma SK, Ranjan R, Kumar V, et al. (2014) Wat1/pop3, a Conserved WD Repeat Containing Protein Acts Synergistically with Checkpoint Kinase Chk1 to Maintain Genome Ploidy in Fission Yeast S. pombe. PLoS ONE 9: e89587.
[23]	Morris GM, Goodsell DS, Halliday RS, et al. (1998) Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function. J Comp Chem 19: 1639-1662.
[24]	Pettersen EF,Goddard TD, Huang CC, et al. (2004) UCSF Chimera—a visualization system for exploratory research and analysis. J Comp Chem 25: 1605-1612.
[25]	Wallace AC, Laskowski RA, Thornton JM (1995) LIGPLOT: a program to generate schematic diagrams of protein-ligand interactions. Protein Eng 8: 127-134.
[26]	Chao WC, Kulkarni K, Zhang Z, et al. (2012) Structure of the mitotic checkpoint complex. Nature 484: 208-213.
[27]	Tange Y, Niwa O (2008) Schizosaccharomyces pombe Bub3 is dispensable for mitotic arrest following perturbed spindle formation. Genetics 179: 785-792.
[28]	Rischitor PE, May KM, Hardwick KG (2007) Bub1 Is a Fission Yeast Kinetochore Scaffold Protein, and Is Sufficient to Recruit other Spindle Checkpoint Proteins to Ectopic Sites on Chromosomes. PLoS ONE 2: e1342. doi: 10.1371/journal.pone.0001342
[29]	He X, Patterson T, Sazer S (1997) The Schizosaccharomyces pombe spindle checkpoint protein mad2p blocks anaphase and genetically interacts with the anaphase-promoting complex. Proc Natl Avad Sci USA 94: 7965-7970. doi: 10.1073/pnas.94.15.7965
[30]	Dorer RK, Zhong S, Tallarico JA, et al. (2005) A small-molecule inhibitor of Mps1 blocks the spindle-checkpoint response to a lack of tension on mitotic chromosomes. Curr Biol 15: 1070-1076.
[31]	Mapelli M, Massimiliano L, Santaguida S, et al. (2007) The Mad2 conformational dimer: structure and implications for the spindle assembly checkpoint, Cell 131: 730-743.
[32]	Izawa D, Pines J (2012) Mad2 and the APC/C compete for the same site on Cdc20 to ensure proper chromosome segregation. J Cell Biol 199: 27-37.
[33]	Acharya BR, Chatterjee A, Ganguli A, et al. (2014) Thymoquinone inhibits microtubule polymerization by tubulin binding and causes mitotic arrest following apoptosis in A549 cells. Biochimie 97: 78-91.
[34]	Huzil J, Chen K, Kurgan L, et al. (2007) The roles of beta-tubulin mutations and isotype expression in acquired drug resistance. Cancer Inform 3: 159-181.

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