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. 1985 Jul 11;13(13):4661–4676. doi: 10.1093/nar/13.13.4661

Characteristics of trypanosome variant antigen genes active in the tsetse fly.

A W Cornelissen, G A Bakkeren, J D Barry, P A Michels, P Borst
PMCID: PMC321818  PMID: 4022771

Abstract

Trypanosoma brucei contains a repertoire of more than 100 different genes for Variant Surface Glycoproteins (VSGs). A small and strain-specific fraction of these genes is expressed in the salivary glands of the tsetse fly (M-genes), giving rise to metacyclic Variable Antigen Types (M-VATs). Antibodies produced in a chronic trypanosome infection initiated by syringe inoculation of bloodstream forms into mammals (i.e. against B-VATs), will react with most of the M-VATs suggesting that these B-VATs express VSG genes that are similar or identical to M-genes. We have cloned DNA complementary to the VSG mRNA of four of such B-VATs and used this to characterize the corresponding VSG genes. In three of the four VATs we find a single VSG gene hybridizing with the cDNA probe and we provide supporting evidence that this gene is expressed as an M-gene. In the bloodstream repertoire these genes appear to be activated by duplicative translocation to another telomere. In all four variants the putative M-genes are telomeric and in the three cases where the location of the genes on chromosome-sized DNA molecules could be determined, the genes were located in large DNA, whereas the majority of the telomeric VSG genes are in chromosomes less than 1000 kb. Our results are best explained by models for M-gene activation involving telomeric expression sites for these genes which are separate from those used by bloodstream forms. The implications of these results for vaccination are discussed.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
  2. Barry J. D., Crowe J. S., Vickerman K. Instability of the Trypanosoma brucei rhodesiense metacyclic variable antigen repertoire. Nature. 1983 Dec 15;306(5944):699–701. doi: 10.1038/306699a0. [DOI] [PubMed] [Google Scholar]
  3. Barry J. D., Crowe J. S., Vickerman K. Neutralization of individual variable antigen types in metacyclic populations of Trypanosoma brucei does not prevent their subsequent expression in mice. Parasitology. 1985 Feb;90(Pt 1):79–88. doi: 10.1017/s0031182000049039. [DOI] [PubMed] [Google Scholar]
  4. Barry J. D., Hajduk S. L., Vickerman K., Le Ray D. Detection of multiple variable antigen types in metacyclic populations of Trypanosoma brucei. Trans R Soc Trop Med Hyg. 1979;73(2):205–208. doi: 10.1016/0035-9203(79)90213-x. [DOI] [PubMed] [Google Scholar]
  5. Bernards A., De Lange T., Michels P. A., Liu A. Y., Huisman M. J., Borst P. Two modes of activation of a single surface antigen gene of Trypanosoma brucei. Cell. 1984 Jan;36(1):163–170. doi: 10.1016/0092-8674(84)90085-0. [DOI] [PubMed] [Google Scholar]
  6. Bernards A., Kooter J. M., Borst P. Structure and transcription of a telomeric surface antigen gene of Trypanosoma brucei. Mol Cell Biol. 1985 Mar;5(3):545–553. doi: 10.1128/mcb.5.3.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bernards A., Michels P. A., Lincke C. R., Borst P. Growth of chromosome ends in multiplying trypanosomes. Nature. 1983 Jun 16;303(5918):592–597. doi: 10.1038/303592a0. [DOI] [PubMed] [Google Scholar]
  8. Bernards A., Van der Ploeg L. H., Frasch A. C., Borst P., Boothroyd J. C., Coleman S., Cross G. A. Activation of trypanosome surface glycoprotein genes involves a duplication-transposition leading to an altered 3' end. Cell. 1981 Dec;27(3 Pt 2):497–505. doi: 10.1016/0092-8674(81)90391-3. [DOI] [PubMed] [Google Scholar]
  9. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Campbell D. A., van Bree M. P., Boothroyd J. C. The 5'-limit of transposition and upstream barren region of a trypanosome VSG gene: tandem 76 base-pair repeats flanking (TAA)90. Nucleic Acids Res. 1984 Mar 26;12(6):2759–2774. doi: 10.1093/nar/12.6.2759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Capbern A., Giroud C., Baltz T., Mattern P. Trypanosoma equiperdum: etude des variations antigéniques au cours de la trypanosomose experimentale du lapin. Exp Parasitol. 1977 Jun;42(1):6–13. doi: 10.1016/0014-4894(77)90055-8. [DOI] [PubMed] [Google Scholar]
  12. Cross G. A. Antigenic variation in trypanosomes. Proc R Soc Lond B Biol Sci. 1978 Jun 5;202(1146):55–72. doi: 10.1098/rspb.1978.0057. [DOI] [PubMed] [Google Scholar]
  13. Cross G. A. Identification, purification and properties of clone-specific glycoprotein antigens constituting the surface coat of Trypanosoma brucei. Parasitology. 1975 Dec;71(3):393–417. doi: 10.1017/s003118200004717x. [DOI] [PubMed] [Google Scholar]
  14. Crowe J. S., Barry J. D., Luckins A. G., Ross C. A., Vickerman K. All metacyclic variable antigen types of Trypanosoma congolense identified using monoclonal antibodies. Nature. 1983 Nov 24;306(5941):389–391. doi: 10.1038/306389a0. [DOI] [PubMed] [Google Scholar]
  15. De Lange T., Borst P. Genomic environment of the expression-linked extra copies of genes for surface antigens of Trypanosoma brucei resembles the end of a chromosome. Nature. 1982 Sep 30;299(5882):451–453. doi: 10.1038/299451a0. [DOI] [PubMed] [Google Scholar]
  16. De Lange T., Kooter J. M., Michels P. A., Borst P. Telomere conversion in trypanosomes. Nucleic Acids Res. 1983 Dec 10;11(23):8149–8165. doi: 10.1093/nar/11.23.8149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Frasch A. C., Borst P., Van den Burg J. Rapid evolution of genes coding for variant surface glycoproteins in trypanosomes. Gene. 1982 Feb;17(2):197–211. doi: 10.1016/0378-1119(82)90073-7. [DOI] [PubMed] [Google Scholar]
  18. Hall T., Esser K. Topologic mapping of protective and nonprotective epitopes on the variant surface glycoprotein of the WRATat 1 clone of Trypanosoma brucei rhodesiense. J Immunol. 1984 Apr;132(4):2059–2063. [PubMed] [Google Scholar]
  19. Hoeijmakers J. H., Borst P., van den Burg J., Weissmann C., Cross G. A. The isolation of plasmids containing DNA complementary to messenger RNA for variant surface glycoproteins of Trypanosoma brucei. Gene. 1980 Mar;8(4):391–417. doi: 10.1016/0378-1119(80)90043-8. [DOI] [PubMed] [Google Scholar]
  20. Horowitz H., Thorburn P., Haber J. E. Rearrangements of highly polymorphic regions near telomeres of Saccharomyces cerevisiae. Mol Cell Biol. 1984 Nov;4(11):2509–2517. doi: 10.1128/mcb.4.11.2509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kosinski R. J. Antigenic variation in trypanosomes: a computer analysis of variant order. Parasitology. 1980 Apr;80(2):343–357. doi: 10.1017/s0031182000000809. [DOI] [PubMed] [Google Scholar]
  22. Laurent M., Pays E., Delinte K., Magnus E., Van Meirvenne N., Steinert M. Evolution of a trypanosome surface antigen gene repertoire linked to non-duplicative gene activation. Nature. 1984 Mar 22;308(5957):370–373. doi: 10.1038/308370a0. [DOI] [PubMed] [Google Scholar]
  23. Laurent M., Pays E., Magnus E., Van Meirvenne N., Matthyssens G., Williams R. O., Steinert M. DNA rearrangements linked to expression of a predominant surface antigen gene of trypanosomes. Nature. 1983 Mar 17;302(5905):263–266. doi: 10.1038/302263a0. [DOI] [PubMed] [Google Scholar]
  24. Lenardo M. J., Rice-Ficht A. C., Kelly G., Esser K. M., Donelson J. E. Characterization of the genes specifying two metacyclic variable antigen types in Trypanosoma brucei rhodesiense. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6642–6646. doi: 10.1073/pnas.81.21.6642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Liu A. Y., Van der Ploeg L. H., Rijsewijk F. A., Borst P. The transposition unit of variant surface glycoprotein gene 118 of Trypanosoma brucei. Presence of repeated elements at its border and absence of promoter-associated sequences. J Mol Biol. 1983 Jun 15;167(1):57–75. doi: 10.1016/s0022-2836(83)80034-5. [DOI] [PubMed] [Google Scholar]
  26. Longacre S., Hibner U., Raibaud A., Eisen H., Baltz T., Giroud C., Baltz D. DNA rearrangements and antigenic variation in Trypanosoma equiperdum: multiple expression-linked sites in independent isolates of trypanosomes expressing the same antigen. Mol Cell Biol. 1983 Mar;3(3):399–409. doi: 10.1128/mcb.3.3.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lumsden W. H., Herbert W. J. Pedigrees of the Edinburgh Trypanosoma (Trypanozoon) antigenic types (ETat). Trans R Soc Trop Med Hyg. 1975;69(2):205–208. doi: 10.1016/0035-9203(75)90156-x. [DOI] [PubMed] [Google Scholar]
  28. Michels P. A., Liu A. Y., Bernards A., Sloof P., Van der Bijl M. M., Schinkel A. H., Menke H. H., Borst P., Veeneman G. H., Tromp M. C. Activation of the genes for variant surface glycoproteins 117 and 118 in Trypanosoma brucei. J Mol Biol. 1983 Jun 5;166(4):537–556. doi: 10.1016/s0022-2836(83)80283-6. [DOI] [PubMed] [Google Scholar]
  29. Michels P. A., Van der Ploeg L. H., Liu A. Y., Borst P. The inactivation and reactivation of an expression-linked gene copy for a variant surface glycoprotein in Trypanosoma brucei. EMBO J. 1984 Jun;3(6):1345–1351. doi: 10.1002/j.1460-2075.1984.tb01975.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Miller E. N., Turner M. J. Analysis of antigenic types appearing in first relapse populations of clones of Trypanosoma brucei. Parasitology. 1981 Feb;82(1):63–80. doi: 10.1017/s0031182000041871. [DOI] [PubMed] [Google Scholar]
  31. Myler P., Nelson R. G., Agabian N., Stuart K. Two mechanisms of expression of a predominant variant antigen gene of Trypanosoma brucei. Nature. 1984 May 17;309(5965):282–284. doi: 10.1038/309282a0. [DOI] [PubMed] [Google Scholar]
  32. Pays E., Delauw M. F., Van Assel S., Laurent M., Vervoort T., Van Meirvenne N., Steinert M. Modifications of a Trypanosoma b. brucei antigen gene repertoire by different DNA recombinational mechanisms. Cell. 1983 Dec;35(3 Pt 2):721–731. doi: 10.1016/0092-8674(83)90105-8. [DOI] [PubMed] [Google Scholar]
  33. Pays E., Laurent M., Delinte K., Van Meirvenne N., Steinert M. Differential size variations between transcriptionally active and inactive telomeres of Trypanosoma brucei. Nucleic Acids Res. 1983 Dec 10;11(23):8137–8147. doi: 10.1093/nar/11.23.8137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pays E., Lheureux M., Steinert M. Analysis of the DNA and RNA changes associated with the expression of isotypic variant-specific antigens of trypanosomes. Nucleic Acids Res. 1981 Sep 11;9(17):4225–4238. doi: 10.1093/nar/9.17.4225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pays E., Lheureux M., Vervoort T., Steinert M. Conservation of a variant-specific surface antigen gene in different trypanosome species and sub-species. Mol Biochem Parasitol. 1981 Dec 31;4(5-6):349–357. doi: 10.1016/0166-6851(81)90066-9. [DOI] [PubMed] [Google Scholar]
  36. Raibaud A., Gaillard C., Longacre S., Hibner U., Buck G., Bernardi G., Eisen H. Genomic environment of variant surface antigen genes of Trypanosoma equiperdum. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4306–4310. doi: 10.1073/pnas.80.14.4306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rifkin M. R. Trypanosoma brucei: some properties of the cytotoxic reaction induced by normal human serum. Exp Parasitol. 1978 Dec;46(2):189–206. doi: 10.1016/0014-4894(78)90131-5. [DOI] [PubMed] [Google Scholar]
  38. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  39. Schwartz D. C., Cantor C. R. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell. 1984 May;37(1):67–75. doi: 10.1016/0092-8674(84)90301-5. [DOI] [PubMed] [Google Scholar]
  40. Seed J. R., Edwards R., Sechelski J. The ecology of antigenic variation. J Protozool. 1984 Feb;31(1):48–53. doi: 10.1111/j.1550-7408.1984.tb04288.x. [DOI] [PubMed] [Google Scholar]
  41. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  42. Van Meirvenne N., Janssens P. G., Magnus E. Antigenic variation in syringe passaged populations of Trypanosoma (Trypanozoon) brucei. 1. Rationalization of the experimental approach. Ann Soc Belg Med Trop. 1975;55(1):1–23. [PubMed] [Google Scholar]
  43. Van Meirvenne N., Janssens P. G., Magnus E., Lumsden W. H., Herbert W. J. Antigenic variation in syringe passaged populations of Trypanosoma (Trypanozoon) brucei. II. Comparative studies on two antigenic-type collections. Ann Soc Belg Med Trop. 1975;55(1):25–30. [PubMed] [Google Scholar]
  44. Van der Ploeg L. H., Cornelissen A. W., Barry J. D., Borst P. Chromosomes of kinetoplastida. EMBO J. 1984 Dec 20;3(13):3109–3115. doi: 10.1002/j.1460-2075.1984.tb02266.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Van der Ploeg L. H., Cornelissen A. W., Michels P. A., Borst P. Chromosome rearrangements in Trypanosoma brucei. Cell. 1984 Nov;39(1):213–221. doi: 10.1016/0092-8674(84)90207-1. [DOI] [PubMed] [Google Scholar]
  46. Van der Ploeg L. H., Liu A. Y., Borst P. Structure of the growing telomeres of Trypanosomes. Cell. 1984 Feb;36(2):459–468. doi: 10.1016/0092-8674(84)90239-3. [DOI] [PubMed] [Google Scholar]
  47. Van der Ploeg L. H., Schwartz D. C., Cantor C. R., Borst P. Antigenic variation in Trypanosoma brucei analyzed by electrophoretic separation of chromosome-sized DNA molecules. Cell. 1984 May;37(1):77–84. doi: 10.1016/0092-8674(84)90302-7. [DOI] [PubMed] [Google Scholar]
  48. Van der Ploeg L. H., Valerio D., De Lange T., Bernards A., Borst P., Grosveld F. G. An analysis of cosmid clones of nuclear DNA from Trypanosoma brucei shows that the genes for variant surface glycoproteins are clustered in the genome. Nucleic Acids Res. 1982 Oct 11;10(19):5905–5923. doi: 10.1093/nar/10.19.5905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Vickerman K. Antigenic variation in trypanosomes. Nature. 1978 Jun 22;273(5664):613–617. doi: 10.1038/273613a0. [DOI] [PubMed] [Google Scholar]
  50. Wieslander L. A simple method to recover intact high molecular weight RNA and DNA after electrophoretic separation in low gelling temperature agarose gels. Anal Biochem. 1979 Oct 1;98(2):305–309. doi: 10.1016/0003-2697(79)90145-3. [DOI] [PubMed] [Google Scholar]
  51. Williams R. O., Young J. R., Majiwa P. A., Doyle J. J., Shapiro S. Z. Analyses of variable antigen gene rearrangements in Trypanosoma brucei. Am J Trop Med Hyg. 1980 Sep;29(5 Suppl):1037–1042. doi: 10.4269/ajtmh.1980.29.1037. [DOI] [PubMed] [Google Scholar]
  52. Yao M. C., Yao C. H. Repeated hexanucleotide C-C-C-C-A-A is present near free ends of macronuclear DNA of Tetrahymena. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7436–7439. doi: 10.1073/pnas.78.12.7436. [DOI] [PMC free article] [PubMed] [Google Scholar]

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