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. 1994 Jun 11;22(11):2102–2108. doi: 10.1093/nar/22.11.2102

Allele-specific expression of a variant-specific surface protein (VSP) of Giardia lamblia.

Y Yang 1, R D Adam 1
PMCID: PMC308127  PMID: 8029018

Abstract

The surfaces of Giardia lamblia trophozoites demonstrate variable expression of a set of cysteine-rich surface proteins, called variant-specific surface proteins (VSP). The cloned Giardia line, WBA6, expresses a 170 kD VSP (VSPA6 or CRP170) which contains approximately 18 to 23 copies of a 65 amino acid repeat. We have cloned the expressed vspA6 gene containing 23 repeats from a genomic library as well as copies of the vspA6 gene with only 8 or 9 repeats from both WBA6 and from WB1269, a cloned line derived from WBA6 which has lost the expressed copy of the gene. The recombinant clones containing the genes with only 8 or 9 repeats have 8 nucleotide substitutions in the coding region. All the recombinant clones map to the same chromosomal location, yet RNA sequencing and comparison with the transcript size indicate that only the clone with 23 repeats contains a gene producing a stable transcript. The most likely interpretation of these data is that G.lamblia trophozoites contain multiple alleles of the vspA6 gene of which only one is expressed.

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  1. Adam R. D., Aggarwal A., Lal A. A., de La Cruz V. F., McCutchan T., Nash T. E. Antigenic variation of a cysteine-rich protein in Giardia lamblia. J Exp Med. 1988 Jan 1;167(1):109–118. doi: 10.1084/jem.167.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adam R. D. Chromosome-size variation in Giardia lamblia: the role of rDNA repeats. Nucleic Acids Res. 1992 Jun 25;20(12):3057–3061. doi: 10.1093/nar/20.12.3057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Adam R. D., Nash T. E., Wellems T. E. Telomeric location of Giardia rDNA genes. Mol Cell Biol. 1991 Jun;11(6):3326–3330. doi: 10.1128/mcb.11.6.3326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Adam R. D., Nash T. E., Wellems T. E. The Giardia lamblia trophozoite contains sets of closely related chromosomes. Nucleic Acids Res. 1988 May 25;16(10):4555–4567. doi: 10.1093/nar/16.10.4555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Adam R. D., Yang Y. M., Nash T. E. The cysteine-rich protein gene family of Giardia lamblia: loss of the CRP170 gene in an antigenic variant. Mol Cell Biol. 1992 Mar;12(3):1194–1201. doi: 10.1128/mcb.12.3.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Aggarwal A., Nash T. E. Comparison of two antigenically distinct Giardia lamblia isolates in gerbils. Am J Trop Med Hyg. 1987 Mar;36(2):325–332. doi: 10.4269/ajtmh.1987.36.325. [DOI] [PubMed] [Google Scholar]
  7. Boothroyd J. C., Wang A., Campbell D. A., Wang C. C. An unusually compact ribosomal DNA repeat in the protozoan Giardia lamblia. Nucleic Acids Res. 1987 May 26;15(10):4065–4084. doi: 10.1093/nar/15.10.4065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Borst P., Greaves D. R. Programmed gene rearrangements altering gene expression. Science. 1987 Feb 6;235(4789):658–667. doi: 10.1126/science.3544215. [DOI] [PubMed] [Google Scholar]
  9. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ey P. L., Khanna K. K., Manning P. A., Mayrhofer G. A gene encoding a 69-kilodalton major surface protein of Giardia intestinalis trophozoites. Mol Biochem Parasitol. 1993 Apr;58(2):247–257. doi: 10.1016/0166-6851(93)90046-z. [DOI] [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  12. Gillin F. D., Hagblom P., Harwood J., Aley S. B., Reiner D. S., McCaffery M., So M., Guiney D. G. Isolation and expression of the gene for a major surface protein of Giardia lamblia. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4463–4467. doi: 10.1073/pnas.87.12.4463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gommers-Ampt J., Lutgerink J., Borst P. A novel DNA nucleotide in Trypanosoma brucei only present in the mammalian phase of the life-cycle. Nucleic Acids Res. 1991 Apr 25;19(8):1745–1751. doi: 10.1093/nar/19.8.1745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Istre G. R., Dunlop T. S., Gaspard G. B., Hopkins R. S. Waterborne giardiasis at a mountain resort: evidence for acquired immunity. Am J Public Health. 1984 Jun;74(6):602–604. doi: 10.2105/ajph.74.6.602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kabnick K. S., Peattie D. A. In situ analyses reveal that the two nuclei of Giardia lamblia are equivalent. J Cell Sci. 1990 Mar;95(Pt 3):353–360. doi: 10.1242/jcs.95.3.353. [DOI] [PubMed] [Google Scholar]
  16. Keister D. B. Axenic culture of Giardia lamblia in TYI-S-33 medium supplemented with bile. Trans R Soc Trop Med Hyg. 1983;77(4):487–488. doi: 10.1016/0035-9203(83)90120-7. [DOI] [PubMed] [Google Scholar]
  17. Le Blancq S. M., Korman S. H., Van der Ploeg L. H. Frequent rearrangements of rRNA-encoding chromosomes in Giardia lamblia. Nucleic Acids Res. 1991 Aug 25;19(16):4405–4412. doi: 10.1093/nar/19.16.4405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Le Blancq S. M., Korman S. H., Van der Ploeg L. H. Spontaneous chromosome rearrangements in the protozoan Giardia lamblia: estimation of mutation rates. Nucleic Acids Res. 1992 Sep 11;20(17):4539–4545. doi: 10.1093/nar/20.17.4539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lyon M. F. Epigenetic inheritance in mammals. Trends Genet. 1993 Apr;9(4):123–128. doi: 10.1016/0168-9525(93)90206-w. [DOI] [PubMed] [Google Scholar]
  20. MacDonald R. J., Swift G. H., Przybyla A. E., Chirgwin J. M. Isolation of RNA using guanidinium salts. Methods Enzymol. 1987;152:219–227. doi: 10.1016/0076-6879(87)52023-7. [DOI] [PubMed] [Google Scholar]
  21. Moore G. T., Cross W. M., McGuire D., Mollohan C. S., Gleason N. N., Healy G. R., Newton L. H. Epidemic giardiasis at a ski resort. N Engl J Med. 1969 Aug 21;281(8):402–407. doi: 10.1056/NEJM196908212810802. [DOI] [PubMed] [Google Scholar]
  22. Mowatt M. R., Aggarwal A., Nash T. E. Carboxy-terminal sequence conservation among variant-specific surface proteins of Giardia lamblia. Mol Biochem Parasitol. 1991 Dec;49(2):215–227. doi: 10.1016/0166-6851(91)90065-e. [DOI] [PubMed] [Google Scholar]
  23. Murtagh J. J., Jr, Mowatt M. R., Lee C. M., Lee F. J., Mishima K., Nash T. E., Moss J., Vaughan M. Guanine nucleotide-binding proteins in the intestinal parasite Giardia lamblia. Isolation of a gene encoding an approximately 20-kDa ADP-ribosylation factor. J Biol Chem. 1992 May 15;267(14):9654–9662. [PubMed] [Google Scholar]
  24. Nash T. E., Aggarwal A., Adam R. D., Conrad J. T., Merritt J. W., Jr Antigenic variation in Giardia lamblia. J Immunol. 1988 Jul 15;141(2):636–641. [PubMed] [Google Scholar]
  25. Nash T. E., Aggarwal A. Cytotoxicity of monoclonal antibodies to a subset of Giardia isolates. J Immunol. 1986 Apr 1;136(7):2628–2632. [PubMed] [Google Scholar]
  26. Nash T. E., Banks S. M., Alling D. W., Merritt J. W., Jr, Conrad J. T. Frequency of variant antigens in Giardia lamblia. Exp Parasitol. 1990 Nov;71(4):415–421. doi: 10.1016/0014-4894(90)90067-m. [DOI] [PubMed] [Google Scholar]
  27. Nash T. E., Herrington D. A., Levine M. M., Conrad J. T., Merritt J. W., Jr Antigenic variation of Giardia lamblia in experimental human infections. J Immunol. 1990 Jun 1;144(11):4362–4369. [PubMed] [Google Scholar]
  28. Nash T. E., Herrington D. A., Losonsky G. A., Levine M. M. Experimental human infections with Giardia lamblia. J Infect Dis. 1987 Dec;156(6):974–984. doi: 10.1093/infdis/156.6.974. [DOI] [PubMed] [Google Scholar]
  29. Nash T. E., Keister D. B. Differences in excretory-secretory products and surface antigens among 19 isolates of Giardia. J Infect Dis. 1985 Dec;152(6):1166–1171. doi: 10.1093/infdis/152.6.1166. [DOI] [PubMed] [Google Scholar]
  30. Nash T. E., McCutchan T., Keister D., Dame J. B., Conrad J. D., Gillin F. D. Restriction-endonuclease analysis of DNA from 15 Giardia isolates obtained from humans and animals. J Infect Dis. 1985 Jul;152(1):64–73. doi: 10.1093/infdis/152.1.64. [DOI] [PubMed] [Google Scholar]
  31. Nash T. E., Merritt J. W., Jr, Conrad J. T. Isolate and epitope variability in susceptibility of Giardia lamblia to intestinal proteases. Infect Immun. 1991 Apr;59(4):1334–1340. doi: 10.1128/iai.59.4.1334-1340.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Orr-Weaver T. L., Szostak J. W. Fungal recombination. Microbiol Rev. 1985 Mar;49(1):33–58. doi: 10.1128/mr.49.1.33-58.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Smith P. D., Gillin F. D., Spira W. M., Nash T. E. Chronic giardiasis: studies on drug sensitivity, toxin production, and host immune response. Gastroenterology. 1982 Oct;83(4):797–803. [PubMed] [Google Scholar]
  35. Van der Ploeg L. H., Gottesdiener K., Lee M. G. Antigenic variation in African trypanosomes. Trends Genet. 1992 Dec;8(12):452–457. doi: 10.1016/0168-9525(92)90330-7. [DOI] [PubMed] [Google Scholar]
  36. Zomerdijk J. C., Kieft R., Duyndam M., Shiels P. G., Borst P. Antigenic variation in Trypanosoma brucei: a telomeric expression site for variant-specific surface glycoprotein genes with novel features. Nucleic Acids Res. 1991 Apr 11;19(7):1359–1368. doi: 10.1093/nar/19.7.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]

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