Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
권호 표지
DOI QR코드

DOI QR Code

Identification of a Novel Microtubule-Binding Protein in Giardia lamblia

  • Kim, Juri (Department of Environmental Medical Biology and Institute of Tropical Medicine, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine) ;
  • Park, Soon-Jung (Department of Environmental Medical Biology and Institute of Tropical Medicine, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine)
  • Received : 2015.12.03
  • Accepted : 2016.05.26
  • Published : 2016.08.31
Download PDF
⟨ Previous Next ⟩

Abstract

Giardia lamblia is a protozoan that causes diarrheal diseases in humans. Cytoskeletal structures of Giardia trophozoites must be finely reorganized during cell division. To identify Giardia proteins which interact with microtubules (MTs), Giardia lysates were incubated with in vitro-polymerized MTs and then precipitated by ultracentifugation. A hypothetical protein (GL50803_8405) was identified in the precipitated fraction with polymerized MTs and was named GlMBP1 (G. lamblia microtubule-binding protein 1). Interaction of GlMBP1 with MTs was confirmed by MT binding assays using recombinant GlMBP1 (rGlMBP1). In vivo expression of GlMBP1 was shown by a real-time PCR and western blot analysis using anti-rGlMBP1 antibodies. Transgenic G. lamblia trophozoites were constructed by integrating a chimeric gene encoding hemagglutinin (HA)-tagged GlMBP1 into a Giardia chromosome. Immunofluorescence assays of this transgenic G. lamblia, using anti-HA antibodies, revealed that GlMBP1 mainly localized at the basal bodies, axonemes, and median bodies of G. lamblia trophozoites. This result indicates that GlMBP1 is a component of the G. lamblia cytoskeleton.

Keywords

References

  1. Elmendorf HG, Dawson SC, McCaffery JM. The cytoskeleton of Giardia lamblia. Int J Parasitol 2003; 33: 3-28. https://doi.org/10.1016/S0020-7519(02)00228-X
  2. Desai A, Mitchison TJ. Microtubule polymerization dynamics. Annu Rev Cell Dev Biol 1997; 13: 83-117. https://doi.org/10.1146/annurev.cellbio.13.1.83
  3. Akhmanova A, Steinmetz MO. Tracking the ends: a dynamic protein network controls the fate of microtubule tips. Nat Rev Mol Cell Biol 2008; 9: 309-322. https://doi.org/10.1038/nrm2369
  4. Dawson SC, Sagolia MS, Mancuso JJ, Woessner DJ, House SA, Friz-Laylin L, Cande WZ. Kinesin-13 regulates flagellar, interphase, mitotic microtubule dynamics in Giardia intestinalis. Eukaryot Cell 2007; 6: 2354-2364. https://doi.org/10.1128/EC.00128-07
  5. Kim J, Nagami S, Lee KH, Park SJ. Characterization of microtubule-binding and dimerization activity of Giardia lamblia end-binding 1 protein. PLoS One 2014; 9: e97850. https://doi.org/10.1371/journal.pone.0097850
  6. Kim J, Sim S, Kim J, Song K, Yong TS, Park SJ. Giardia lamblia EB1 is a functional homolog of yeast Bim1p that binds to microtubules. Parasitol Int 2008; 57: 465-471. https://doi.org/10.1016/j.parint.2008.05.008
  7. Holberton DV, Ward AP. Isolation of the cytoskeleton from Giardia. Tubulin and a low-molecular-weight protein associated with microribbon structures. J Cell Sci 1981; 47: 39-166.
  8. Lourenco D, Andrade Ida S, Terra LL, Guimaraes PR, Zingali RB, de Souza W. Proteomic analysis of the ventral disc of Giardia lamblia. BMC Res Notes 2012; 5: 41. https://doi.org/10.1186/1756-0500-5-41
  9. Hagen KD, Hirakawa MP, House SA, Schwartz CL, Pham JK, Cipriano MJ, De La Torre MJ, Sek AC, Du G, Forsythe BM, Dawson SC. Novel structural components of the ventral disc and lateral crest in Giardia intestinalis. PLoS Negl Trop Dis 2011; 5: e1442. https://doi.org/10.1371/journal.pntd.0001442
  10. Woessner DJ, Dawson SC. The Giardia median body protein is a ventral disc protein that is critical for maintaining a domed disc conformation during attachment. Eukaryot Cell 2012; 11: 292-301. https://doi.org/10.1128/EC.05262-11
  11. Keister DB. Axenic culture of Giardia lamblia in TYI-S-33 medium supplemented with bile. Trans R Soc Trop Med Hyg 1983; 77: 487-488. https://doi.org/10.1016/0035-9203(83)90120-7
  12. Harlow E, Lane D. Immunizations. In Antibodies: A Laboratory Manual, Immunization. New York, USA. Cold Spring Harbor Laboratory. 1988. pp 53-63.
  13. Singer SM, Yee J, Nash TE. Episomal and integrated maintenance of foreign DNA in Giardia lamblia. Mol Biochem Parasitol 1998; 92: 59-69. https://doi.org/10.1016/S0166-6851(97)00225-9
  14. Kim J, Bae S, Sung M, Lee K, Park SJ. Comparative proteomic analysis of trophozoites versus cysts of Giardia lamblia. Parasitol Res 2009; 104: 475-479. https://doi.org/10.1007/s00436-008-1223-x
  15. Gourguechon S, Cande WZ. Rapid tagging and integration of genes in Giardia intestinalis. Eukaryot Cell 2011; 10: 142-145. https://doi.org/10.1128/EC.00190-10
  16. Debec A, Sullivan W, Bettencourt-Diaz M. Centrioles: active players or passengers during mitosis? Cell Mol Life Sci 2010; 67: 21173-2194.
  17. Crossley R, Marshall J, Cark JT, Holberton DV. Immunocytochemical differentiation of microtubules in the cytoskeleton of Giardia lamblia using monoclonal antibodies to $\gamma$-tubulin and polyclonal antibodies to associated low molecular weight proteins. J Cell Sci 1986; 80: 233-252.
  18. Nohynkova E, Draber P, Reischig J, Kulda J. Localization of gamma-tubuin in interphase and mitotic cells of a unicellular eukaryote, Giardia intestinalis. Eur J Cell Biol 2000; 79: 438-445. https://doi.org/10.1078/0171-9335-00066
  19. Lauwaet T, Smith AJ, Reiner DS, Romijn EP, Wong CCL, Davids BJ, Shah SA, Yates JR, Gillin FD. Mining the Giardia genome and proteome for conserved and unique basal body proteins. Int J Parasitol 2011; 41: 1079-1092. https://doi.org/10.1016/j.ijpara.2011.06.001

Cited by

  1. Giardia intestinalis coiled-coil cytolinker protein 259 interacts with actin and tubulin vol.120, pp.3, 2016, https://doi.org/10.1007/s00436-021-07062-6