Abstract
The proteome of Giardia lamblia at its cyst stage was compared with that of trophozoites by using two-dimensional SDS-PAGE gel electrophoresis. Protein spots that increased in the extracts of cysts compared to trophozoites were identified by MALDI-TOF mass spectroscopy and categorized as cytoskeletal proteins, metabolic enzymes, a cell-cycle-specific kinase, stress resistance proteins, and a protein involved in translation. Expression patterns of five of the identified proteins were examined during encystation by real-time PCR. Expression of cwp1 (encoding cyst wall protein 1), a marker for encystation, was increased 11-fold. In contrast, tim (encoding triose-1-phosphate isomerase) was expressed constitutively during encystation, and its transcription level was therefore used as a mRNA loading control. Expression of three genes encoding β-tubulin, vacuolar ATPase, and never-in-mitosis-A-related protein kinase did not vary significantly during encystation. Interestingly, genes encoding two heat shock proteins (Hsp70 and Hsp90) showed increased expression during encystation, suggesting that this differentiation process accompanies a cellular response to stress in G. lamblia.
References
Adam RD (1991) The biology of Giardia spp. Microbiol Rev 55:706–732
Best AA, Morrison HG, McArthur AG, Sogin ML, Olsen GJ (2004) Evolution of eukaryotic transcription: insights from the genome of Giardia lamblia. Genome Res 14:1537–1547
Bradley BA, Quarmby LM (2005) A NIMA-related kinase, Cnk2p, regulates both flagellar length and cell size in Chlamydomonas. J Cell Sci 118:3317–3326
Elmendorf HG, Singer SM, Pierce J, Cowan J, Nash TE (2001) Initiator and upstream elements in the a2-tubulin promoter of Giardia lamblia. Mol Biochem Parasitol 113:157–169
Gillin FD, Reiner DS (1996) Cell biology of the primitive eukaryote Giardia lamblia. Annu Rev Microbiol 50:679–705
Holberton DV, Marshall J (1995) Analysis of consensus sequence patterns in Giardia cytoskeleton gene promoters. Nucleic Acids Res 23:2945–2953
Kane AV, Ward HD, Keusch GT, Pereira MEA (1991) In vitro encystation of Giardia lamblia: Large-scale production of in vitro cysts and strain and clone differences in encystation efficiency. J Parasitol 77:974–981
Keister DB (1983) Axenic culture of Giardia lamblia in TYI-S-33 medium supplemented with bile. Trans R Soc Trop Med Hyg 77:487–488
Knodler LA, Svard SG, Silberman JD, Davids BJ, Gillin FD (1999) Developmental gene regulation in Giardia lamblia: first evidence for an encystation-specific promoter and differential 5′ mRNA processing. Mol Microbiol 34:327–340
Lauweat T, Davids BJ, Reiner DS, Gillin F (2007) Encystation of Giardia lamblia: a model for other parasites. Curr Opin Microbiol 10:554–559
Lujan HD, Mowatt MR, Conrad JT, Bowers B, Nash TE (1995) Identification of a novel Giardia lamblia cyst wall protein with leucine-rich repeats. Implications for secretory granule formation and protein assembly into the cyst wall. J Biol Chem 270:29307–29313
Lujan HD, Mowatt MR, Conrad JT, Nash TE (1996) Increased expression of the molecular chaperon BiP/GRP78 during differentiation of a primitive eukaryote. Biol Cell 86:11–18
Mowatt MR, Lujan HD, Cotton DB, Boweers B, Yee J, Nash TE (1995) Developmentally regulated expression of a Giardia lamblia cyst wall protein gene. Mol Microbiol 15:955–963
Que X, Svard SG, Meng T, Hetsko ML, Aley SB, Gillin FD (1996) Developmentally regulated transcripts and evidence of differential mRNA processing in Giardia lamblia. Mol Biochem Parasitol 81:101–110
Reiner DS, Shinnick TM, Ardeshir F, Gillin FD (1992) Encystation of Giardia lamblia leads to expression of antigens recognized by antibodies against conserved heat shock proteins. Infect Immun 60:5312–5315
Wandinger SK, Richter K, Buchner J (2008) The Hsp90 chaperon machinery. J Biol Chem 283:18473–18477
Wiech H, Buchner J, Zimmermann R, Jakob U (1992) Hsp90 chaperones protein folding in vitro. Nature 358:169–170
Yang HW, Chung HJ, Yong TS, Lee BH, Park SJ (2003) Identification of an encystations-specific transcription factor, Myb protein in Giardia lamblia. Mol Biochem Parasitol 128:167–174
Yee J, Mowatt MR, Dennis PP, Nash TE (2000) Transcriptional analysis of the glutamate dehydrogenase gene in the primitive eukaryote, Giardia lamblia. Identification of a primordial gene promoter. J Biol Chem 275:11432–11439
Acknowledgment
This work was supported by the Seoul R & B D program (grant 10580), South Korea.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kim, J., Bae, SS., Sung, MH. et al. Comparative proteomic analysis of trophozoites versus cysts of Giardia lamblia . Parasitol Res 104, 475–479 (2009). https://doi.org/10.1007/s00436-008-1223-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-008-1223-x