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A Giardia duodenalis gene* encoding a protein with multiple repeats of a toxin homologue

Published online by Cambridge University Press:  06 April 2009

N. Chen ,
J. A. Upcrofta  and
P. Upcroft
N. Chen
Affiliation:
Queensland Institute of Medical Research, 300 Herston Road, Brisbane, Queensland 4029, Australia
J. A. Upcrofta
Affiliation:
Queensland Institute of Medical Research, 300 Herston Road, Brisbane, Queensland 4029, Australia
P. Upcroft
Affiliation:
Queensland Institute of Medical Research, 300 Herston Road, Brisbane, Queensland 4029, Australia
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Summary

A gene, CRP136, from the intestinal protozoan parasite Giardia duodenalis, expressed at a high level in a metronidazole-resistant line, encodes a 136 kDa protein with 23 copies of a 40 amino acid repeat. The protein is cysteine-rich and has the typical membrane-spanning region and CXXC amino acid motifs of a family of Giardia cysteine-rich surface proteins (CRSPs). The repeat unit in CRP136, shares 57% homology with the gene encoding the precursor of the sarafotoxins, a group of snake toxins from the burrowing adder known to cause symptoms similar to those of humans acutely infected with Giardia. The sarafotoxins are low molecular weight sulphydryl cross-linked peptides which are proteolytically cleaved from a precursor polyprotein. CRP136 has homology over the entire length of the sarafotoxin precursor, and the repeats are of the same length. Thus CRP136 represents the first evidence for a potential Giardia toxin. The genomic copy number of CRP136 appears to be the same in both the parent and drug-resistant lines and expression of this gene, and at least one other, is associated with a conserved partial duplication, but not amplification, of one chromosome.

Keywords

toxinGiardia duodenalis, cysteine-rich proteinsarafotoxindrug resistance
Type
Research Article
Information
Parasitology , Volume 111 , Issue 4 , November 1995 , pp. 423 - 431
Copyright
Copyright © Cambridge University Press 1995

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References

REFERENCES

Adam, R. D. (1991). The biology of Giardia spp. Microbiological Reviews 55, 706–32.CrossRefGoogle ScholarPubMed
Adam, R. D., Yang, Y. M. & Nash, T. E. (1992). The cysteine-rich protein gene family of Giardia lamblia: loss of the CRP170 gene in an antigenic variant. Molecular and Cellular Biology 12, 1194–201.Google Scholar
Aggarwal, A., Merritt, J. W. Jr., & Nash, T. E. (1989). Cysteine-rich variant surface proteins of Giardia lamblia. Molecular and Biochemical Parasitology 32, 39–48.CrossRefGoogle ScholarPubMed
Aley, S. B. & Gillin, F. D. (1993). Giardia lamblia: Posttranslational processing and status of exposed cysteine residues in TSA417, a variable surface antigen. Experimental Parasitology 77, 295–305.CrossRefGoogle ScholarPubMed
Bdolah, A., Wollberg, Z. & Kochva, E. (1991). Sarafotoxins: a new group of cardiotoxic peptides from the venom of Atractaspis. In Snake Toxins (ed. Harvey, A. L.), pp. 415–24. New York: Pergamon Press.Google Scholar
Boreham, P. F. L., Phillips, R. E. & Shepherd, R. W. (1986). Activity of drugs against Giardia intestinalis in neonatal mice. Journal of Antimicrobial Chemotherapy 18, 393–8.CrossRefGoogle ScholarPubMed
Chen, N. & Crane, D. I. (1992). Induction of the major integral membrane protein of mouse liver peroxisomes by peroxisome proliferators. The Biochemical Journal 283, 605–10.CrossRefGoogle ScholarPubMed
Chen, N., Upcroft, J. A. & Upcroft, P. (1994). Physical map of a 2 Mb chromosome of the intestinal protozoan parasite Giardia duodenalis. Chromosome Research 2, 307–13.Google ScholarPubMed
Church, G. M. & Gilbert, W. (1984). Genomic sequencing. Proceedings of the National Academy of Sciences, USA 81, 1991–5.CrossRefGoogle ScholarPubMed
De Nucci, G., Thomas, R., D'Orleans-Juste, P., Antunes, E., Walder, C., Warner, T. D. & Vane, J. R. (1988). Pressor effects of circulating endothelin are limited by its removal in pulmonary circulation and by the release of prostacyclin and endothelin-derived relaxing factor. Proceedings of the National Academy of Sciences, USA 85, 9797–800.Google Scholar
Devereux, J., Haeberli, P. & Smithies, O. (1984). A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Research 12, 387–95.CrossRefGoogle ScholarPubMed
Ducancel, F., Matre, V., Dupont, C., Lajeunesse, E., Wollberg, Z., Bdolah, A., Kochva, E., Boulain, J. C. & Menez, A. (1993). Cloning and sequencing analysis of cDNAs encoding presursors of sarafotoxins. Journal of Biological Chemistry 268, 3052–5.CrossRefGoogle Scholar
Feinberg, A. P. & VoGelstein, B. (1984). Addendum. ‘A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity.’ Analytical Biochemistry 137, 266–7.Google ScholarPubMed
Forney, J. D., Epstein, L. M., Preer, L. B., Rudman, B. M., Widmayer, D. J., Klein, W. H. & Preer, J. R. Jr., (1983). Structure and expression of genes for surface proteins in Paramecium. Molecular and Cellular Biology 3, 466–74.Google ScholarPubMed
Gillin, F. D., Hagblom, P., Harwood, J., Aley, S. B., Reiner, D. S., McCaffery, M., So, M. & Guiney, D. G. (1990). Isolation and expression of the gene for a major surface protein of Giardia lamblia. Proceedings of the National Academy of Sciences, USA 87, 4463–7.CrossRefGoogle Scholar
Han, J. H., Law, S. W., Keller, P. M., Kniskern, P. J., Silbergklang, M., Tung, J. S., Gasic, T. B., Friedman, P. A. & Ellis, R. W. (1989). Cloning and expression of cDNA encoding antistasin, a leech-derived protein having anti-coagulant and anti-metastatic properties. Gene 75, 42–57.CrossRefGoogle ScholarPubMed
Hanahan, D. (1983). Studies on transformation of Escherichia coli with plasmids. Journal of Molecular Biology 166, 557–80.CrossRefGoogle ScholarPubMed
Hanahan, D. & Meselson, M. (1980). Plasmid screening in high colony density. Gene 10, 63–7.CrossRefGoogle ScholarPubMed
Holmgren, J. (1981). Actions of cholera toxin and the prevention of cholera. Nature, London 292, 413–17.Google ScholarPubMed
Inoue, A., Yanagisawa, M., Kimura, S., Kasuya, Y., Miyauchi, T., Goto, K. & Masaki, T. (1989). The human endothelin family: three structurally and pharmacologically distinct isopeptides predicted by three separate genes. Proceedings of the National Academy of Sciences, USA 86, 2863–7.CrossRefGoogle ScholarPubMed
Keusch, G. Y., Donohue-Rolfe, A. & Jacewicz, M. (1982). Shigella toxin(s): description and role in diarrhea and dysentery. Pharmacological Therapeutics 15, 403–38.CrossRefGoogle Scholar
Laskowski, M. Jr. & Kato, I. (1980). Protein inhibitors of proteinases. Annual Review of Biochemistry 49, 593–626.CrossRefGoogle ScholarPubMed
Lin, H., Lei, S. & Wilcox, G. (1985). An improved DNA sequencing strategy. Analytical Biochemistry 147, 114–19.Google ScholarPubMed
Lipman, D. J. & Pearson, W. R. (1985). Rapid and sensitive protein similarity searches. Science 227 1435–41.CrossRefGoogle ScholarPubMed
Lohman, K. L., Langer, P. L. & Mcmahon-Pratt, D. (1990). Molecular cloning and characterization of the immunologically protective surface glycoprotein GP46/M-2 of Leishmania amazonensis. Proceedings of the National Academy of Sciences, USA 87, 8393–7.CrossRefGoogle ScholarPubMed
Maniatis, T., Fritsch, E. F. & Sambrook, J. (1982). Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press.Google Scholar
Mann, B. J. & Petri, W. A. Jr., (1991). Cell surface proteins of Entamoeba histolytica. Parasitology Today 7, 173–6.CrossRefGoogle ScholarPubMed
Nash, T. E., Aggarwal, A., Adam, R. D., Conrad, J. T. & Merritt, J. W. Jr., (1988). Antigenic variation in Giardia lamblia. Journal of Immunology 141, 636–41.CrossRefGoogle ScholarPubMed
Nash, T. E. & Keister, D. B. (1985). Differences in excretory-secretory products and surface antigens among 19 isolates of Giardia. Journal of Infectious Diseases 152, 1166–71.CrossRefGoogle Scholar
Nash, T. E. & Mowatt, M. R. (1992). Characterization of a Giardia lamblia variant-specific surface protein (VSP) gene from isolate GS/M and estimation of the VSP gene repertoire size. Molecular and Biochemical Parasitology 51, 219–28.CrossRefGoogle ScholarPubMed
Nash, T. E. & Mowatt, M. R. (1993). Variant-specific surface proteins of Giardia lamblia are zinc-binding proteins. Proceedings of the National Academy of Sciences, USA 90, 5489–93.Google ScholarPubMed
Prat, A. (1990). Conserved sequences flank variable tandem repeats in two alleles of the G protein of Paramecium primaurelia. Journal of Molecular Biology 211, 521–35.CrossRefGoogle Scholar
Sanger, F., Nicklen, S. & Coulson, A. R. (1977). DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, USA 74, 5463–7.CrossRefGoogle ScholarPubMed
Smith, T. F. & Waterman, M. S. (1981). Comparison of biosequences. Advances in Applied Mathematics 2, 482–9.CrossRefGoogle Scholar
Southern, E. M. (1975). Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98, 503–17.CrossRefGoogle ScholarPubMed
Townson, S. M., Laqua, H., Upcroft, P., Boreham, P. F. L. & Upcroft, J. A. (1992). Induction of metronidazole and furazolidone resistance in Giardia. Transactions of the Royal Society for Tropical Medicine and Hygiene 86, 521–2.CrossRefGoogle ScholarPubMed
Trezise, A. E., Godfrey, E. A., Holmes, R. S. & Beacham, I. R. (1989). Cloning and sequencing of cDNA encoding baboon liver alcohol dehydrogenase: evidence for a common ancestral lineage with the human alcohol dehydrogenase β subunit and for class I ADH gene duplications predating primate radiation. Proceedings of the National Academy of Sciences, USA 86, 5454–8.Google Scholar
Upcroft, J. A., Boreham, P. F. L. & Upcroft, P. (1989). Geographic variation in Giardia karyotypes. International Journal for Parasitology 19, 519–27.Google ScholarPubMed
Upcroft, J. A., Capon, A. G., Dharmkrong-At, A., Healey, A., Boreham, P. F. L. & Upcroft, P. (1987). Giardia intestinalis antigens expressed in Escherichia coli. Molecular and Biochemical Parasitology 26, 267–76.CrossRefGoogle ScholarPubMed
Upcroft, J. A., Healey, A. & Upcroft, P. (1993). Chromosomal duplication in Giardia intestinalis. International Journal for Parasitology 23, 609–16.CrossRefGoogle Scholar
Upcroft, P. & Healey, A. (1987). Rapid and efficient method for cloning of blunt-ended DNA fragments. Gene 51, 69–75.Google ScholarPubMed
Wolfe, M. S. (1992). Giardiasis. Clinical Microbiology Reviews 5, 93–100.CrossRefGoogle ScholarPubMed
World Health Organization (1990). Drugs Used in Parasitic Diseases. Geneva: World Health Organization.Google Scholar
Yanagisawa, M., Kurihara, H., Kimura, S., Tomobe, Y., Kobayashi, M., Mitsui, Y., Yazaki, Y., Goto, K. & Masaki, T. (1988). A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature, London 332, 411–15.CrossRefGoogle ScholarPubMed
Zasloff, M. (1987). Magainins, a class of antimicrobial peptides from Xenopus skin: Isolation, characterization of two active forms, and partial cDNA sequence of a precursor. Proceedings of the National Academy of Sciences, USA 84, 5449–53.CrossRefGoogle ScholarPubMed