Hostname: page-component-78c5997874-j824f Total loading time: 0 Render date: 2024-11-17T19:12:09.075Z Has data issue: false hasContentIssue false

Molecular and biochemical mining of heat-shock and 14-3-3 proteins in drug-induced protoscolices of Echinococcus granulosus and the detection of a candidate gene for anthelmintic resistance

Published online by Cambridge University Press:  25 August 2010

D. Pan
Affiliation:
Indian Veterinary Research Institute, Eastern Regional Station, 37-Belgachia Road, Kolkata700 037, India
S. Das
Affiliation:
Indian Veterinary Research Institute, Eastern Regional Station, 37-Belgachia Road, Kolkata700 037, India
A.K. Bera
Affiliation:
Indian Veterinary Research Institute, Eastern Regional Station, 37-Belgachia Road, Kolkata700 037, India
S. Bandyopadhyay
Affiliation:
Indian Veterinary Research Institute, Eastern Regional Station, 37-Belgachia Road, Kolkata700 037, India
S. Bandyopadhyay
Affiliation:
NRC on Yak, Arunachal Pradesh, India
S. De
Affiliation:
Indian Veterinary Research Institute, Eastern Regional Station, 37-Belgachia Road, Kolkata700 037, India
T. Rana
Affiliation:
Indian Veterinary Research Institute, Eastern Regional Station, 37-Belgachia Road, Kolkata700 037, India
S.K. Das
Affiliation:
Indian Veterinary Research Institute, Eastern Regional Station, 37-Belgachia Road, Kolkata700 037, India
V.V. Suryanaryana
Affiliation:
Indian Veterinary Research Institute, FMD Research Laboratory, Bangalore Campus, Hebbal, Bangalore560 024, India
J. Deb
Affiliation:
Department of Chest Medicine, RG Kar Medical College and Hospital, Kolkata, West Bengal, India
D. Bhattacharya*
Affiliation:
Indian Veterinary Research Institute, Eastern Regional Station, 37-Belgachia Road, Kolkata700 037, India
*
*Fax: +913325565725 E-mail: [email protected]

Abstract

Cystic echinococcosis (CE) caused by the larval stage of Echinococcus granulosus is a disease that affects both humans and animals. In humans the disease is treated by surgery with a supplementary option of chemotherapy with a benzimidazole compound. During the present study heat-shock protein 60 (HSP 60) was identified as one of the most frequently expressed biomolecules by E. granulosus after albendazole treatment. Data were correlated with 14-3-3 protein signature, and overexpression of this molecule after albendazole induction was an indicator of cell survival and signal transduction during in vitro maintenance of E. granulosus for up to 72 h. This observation was further correlated with a uniform expression pattern of a housekeeping gene (actin II). Out of three β-tubulin gene isoforms of E. granulosus, β-tubulin gene isoform 2 showed a conserved point mutation indicative of benzimidazole resistance.

Type
Regular research papers
Copyright
Copyright © Cambridge University Press 2010

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Ammann, R. & Eckert, J. (1995) Clinical diagnosis and treatment of echinococcosis in humans. pp. 411463in Thompson, R.C.A. & Lymbery, A.J. (Eds) Echinococcus and hydatid disease. Wallingford, UK, CAB International.Google Scholar
Brehm, K., Kronthaler, K., Jura, H. & Frosch, M. (2000) Cloning and characterization of beta-tubulin genes from Echinococcus multilocularis. Molecular and Biochemical Parasitology 107, 297302.Google Scholar
Casado, N., Rodriguez-Caabeiro, F. & Fernandez, S. (1986) In vitro survival of Echinococcus granulosus protoscolices in several media, at +4 degrees C and +37 degrees C. Zeitschrift für Parasitenkunde 72, 273278.CrossRefGoogle Scholar
Del Razo, L.M., Quintanilla-Vega, B., Brambila-Colombres, E., Calderón-Aranda, E.S., Manno, M. & Albores, A. (2001) Stress proteins induced by arsenic. Toxicology and Applied Pharmacology 177, 132148.CrossRefGoogle ScholarPubMed
Driscoll, M., Dean, E., Reilly, E., Bergholz, E. & Chalfie, M. (1989) Genetic and molecular analysis of a Caenorhabditis elegans beta-tubulin that conveys benzimidazole sensitivity. Journal of Cell Biology 109, 29933003.Google Scholar
Elard, L., Comes, A.M. & Humbert, J.F. (1996) Sequences of beta-tubulin cDNA from benzimidazole-susceptible and -resistant strains of Teladorsagia circumcincta, a nematode parasite of small ruminants. Molecular and Biochemical Parasitology 79, 249253.CrossRefGoogle ScholarPubMed
Feder, M.E. & Hofmann, G.E. (1999) Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annual Review of Physiology 61, 243282.Google Scholar
Fu, H., Subramanian, R.R. & Masters, S.C. (2000) 14-3-3 proteins: structure, function, and regulation. Annual Review of Pharmacology and Toxicology 40, 617647.Google Scholar
Geerts, S. & Gryseels, B. (2000) Drug resistance in human helminths: current situation and lessons from livestock. Clinical Microbiology 13, 207222.CrossRefGoogle ScholarPubMed
Jung, M.K. & Oakley, B.R. (1990) Identification of an amino acid substitution in the benA, beta-tubulin gene of Aspergillus nidulans that confers thiabendazole resistance and benomyl supersensitivity. Cell Motility and Cytoskeleton 17, 8794.CrossRefGoogle ScholarPubMed
Kaufmann, S.H. (1992) The cellular immune response to heat shock proteins. Experientia 48, 640643.CrossRefGoogle ScholarPubMed
Kwa, M.S., Veenstra, J.G. & Roos, M.H. (1993a) Benzimidazole resistance in Haemonchus contortus is correlated with a conserved mutation at amino acid 200 in beta-tubulin isotype 1. Molecular and Biochemical Parasitology 63, 299303.CrossRefGoogle Scholar
Kwa, M.S., Veenstra, J.G. & Roos, M.H. (1993b) Molecular characterisation of beta-tubulin genes present in benzimidazole-resistant populations of Haemonchus contortus. Molecular and Biochemical Parasitology 60, 133143.CrossRefGoogle ScholarPubMed
Kwa, M.S., Veenstra, J.G., Van Dijk, M. & Roos, M.H. (1995) Beta-tubulin genes from the parasitic nematode Haemonchus contortus modulate drug resistance in Caenorhabditis elegans. Journal of Molecular Biology 246, 500510.CrossRefGoogle ScholarPubMed
Maresca, B. & Kobayashi, G.S. (1994) Hsp70 in parasites: as an inducible protective protein and as an antigen. Experientia 50, 10671074.CrossRefGoogle ScholarPubMed
Martínez, J., Pérez-Serrano, J., Bernadina, W.E. & Rodriguez-Caabeiro, F. (1999a) Echinococcus granulosus: in vitro effects of ivermectin and praziquantel on hsp60 and hsp70 levels. Experimental Parasitology 93, 171180.Google Scholar
Martínez, J., Pérez-Serrano, J., Bodega, G., Casado, N. & Rodríguez-Caabeiro, F. (1999b) Heat shock proteins HSP70 and HSP60 in Echinococcus granulosus protoscolices. Folia Parasitologica (Praha) 46, 7678.Google ScholarPubMed
Matsumoto, J., Müller, N., Hemphill, A., Oku, Y., Kamiya, M. & Gottstein, B. (2006) 14-3-3 and II/3-10-gene expression as molecular markers to address viability and growth activity of Echinococcus multilocularis metacestodes. Parasitology 132, 8394.Google Scholar
Polla, B.S. (1991) Heat shock proteins in host–parasite interactions. Immunology Today 12, 3841.CrossRefGoogle ScholarPubMed
Roos, M.H., Boersema, J.H., Borgsteede, F.H., Cornelissen, J., Taylor, M. & Ruitenberg, E.J. (1990) Molecular analysis of selection for benzimidazole resistance in the sheep parasite Haemonchus contortus. Molecular and Biochemical Parasitology 43, 7788.CrossRefGoogle ScholarPubMed
Roos, M.H., Kwa, M.S.G. & Grant, W.N. (1995) New genetic and practical implications of selection for anthelmintic resistance in parasitic nematodes. Parasitology Today 11, 148150.CrossRefGoogle Scholar
Siles-Lucas, M., Merli, M. & Gottstein, B. (2008) 14-3-3 proteins in Echinococcus: their role and potential as protective antigens. Experimental Parasitology 119, 516523.CrossRefGoogle ScholarPubMed
Sullivan, K.F. (1988) Structure and utilization of tubulin isotypes. Annual Review of Cell Biology 4, 687716.CrossRefGoogle ScholarPubMed
Tamura, K., Dudley, J., Nei, M. & Kumar, S. (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24, 15961599.Google Scholar
Thomas, J.H., Neff, N.F. & Botstein, D. (1995) Isolation and characterization of mutations in the beta-tubulin gene of Saccharomyces cerevisiae. Genetics 111, 715734.CrossRefGoogle Scholar
Towbin, H., Staehelin, T. & Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Science, USA 76, 43504354.Google Scholar
van Hemert, M.J., Steensma, H.Y. & van Heusden, G.P. (2001) 14-3-3 proteins: key regulators of cell division, signalling and apoptosis. Bioessays 23, 936946.CrossRefGoogle ScholarPubMed
van Heusden, G. & Steensma, H.Y. (2006) Yeast 14-3-3 proteins. Yeast 23, 159171.Google Scholar
Xie, T., Ho, S.L. & Ma, O.C. (1997) High resolution single strand conformation polymorphism analysis using formamide and ethidium bromide staining. Molecular Pathology 50, 276278.CrossRefGoogle ScholarPubMed