Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1983;2(10):1721–1728. doi: 10.1002/j.1460-2075.1983.tb01648.x

Repression of glycoprotein synthesis and release of surface coat during transformation of Trypanosoma brucei.

P Overath, J Czichos, U Stock, C Nonnengaesser
PMCID: PMC555349  PMID: 6688990

Abstract

The biosynthesis of the variant surface glycoprotein (VSG) and its release from the surface of Trypanosoma brucei 427 variant clone MITat 1.4 (117) during in vitro transformation of bloodstream trypomastigotes to procyclic trypomastigotes was investigated. After transfer to the transformation medium at 27 degrees C, VSG synthesis is repressed with a half-time, t1/2 = 30 min. Concomitantly VSG-specific mRNA is lost suggesting that repression operates at the transcriptional level. The expression-linked extra gene copy, which codes for VSG, is retained during and after completion of transformation. After repression of VSG synthesis, surface VSG is shed from the cells into the culture medium. During release part of VSG (apparent mol. wt. 61 000) is proteolytically cleaved to a product (apparent mol. wt. 51 000) which represents the N-terminal domain of the protein as judged by the absence of the carbohydrate moiety normally linked to the C terminus.

Full text

PDF
1721

Images in this article

Selected References

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

  1. Allen G., Gurnett L. P., Cross G. A. Complete amino acids sequence of a variant surface glycoprotein (VSG 117) from Trypanosoma brucei. J Mol Biol. 1982 May 25;157(3):527–546. doi: 10.1016/0022-2836(82)90474-0. [DOI] [PubMed] [Google Scholar]
  2. Baltz T., Giroud C., Baltz D., Duvillier G., Degand P., Demaille J., Pautrizel R. The variable surface glycoproteins of Trypanosoma equiperdum are phosphorylated. EMBO J. 1982;1(11):1393–1398. doi: 10.1002/j.1460-2075.1982.tb01328.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barry J. D., Vickerman K. Trypanosoma brucei: loss of variable antigens during transformation from bloodstream to procyclic forms in vitro. Exp Parasitol. 1979 Oct;48(2):313–324. doi: 10.1016/0014-4894(79)90114-0. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Bienen E. J., Hammadi E., Hill G. C. Initiation of trypanosome transformation from bloodstream trypomastigotes to procyclic trypomastigotes. J Parasitol. 1980 Aug;66(4):680–682. [PubMed] [Google Scholar]
  6. Bienen E. J., Hammadi E., Hill G. C. Trypanosoma brucei: biochemical and morphological changes during in vitro transformation of bloodstream- to procyclic-trypomastigotes. Exp Parasitol. 1981 Jun;51(3):408–417. doi: 10.1016/0014-4894(81)90128-4. [DOI] [PubMed] [Google Scholar]
  7. Bienen E. J., Hill G. C., Shin K. O. Elaboration of mitochondrial function during Trypanosoma brucei differentiation. Mol Biochem Parasitol. 1983 Jan;7(1):75–86. doi: 10.1016/0166-6851(83)90118-4. [DOI] [PubMed] [Google Scholar]
  8. Boothroyd J. C., Cross G. A. Transcripts coding for variant surface glycoproteins of Trypanosoma brucei have a short, identical exon at their 5' end. Gene. 1982 Dec;20(2):281–289. doi: 10.1016/0378-1119(82)90046-4. [DOI] [PubMed] [Google Scholar]
  9. Boothroyd J. C., Paynter C. A., Coleman S. L., Cross G. A. Complete nucleotide sequence of complementary DNA coding for a variant surface glycoprotein from Trypanosoma brucei. J Mol Biol. 1982 May 25;157(3):547–556. doi: 10.1016/0022-2836(82)90475-2. [DOI] [PubMed] [Google Scholar]
  10. Borst P., Cross G. A. Molecular basis for trypanosome antigenic variation. Cell. 1982 Jun;29(2):291–303. doi: 10.1016/0092-8674(82)90146-5. [DOI] [PubMed] [Google Scholar]
  11. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  12. Brown R. C., Evans D. A., Vickerman K. Changes in oxidative metabolism and ultrastructure accompanying differentiation of the mitochondrion in Trypanosoma brucei. Int J Parasitol. 1973 Sep;3(5):691–704. doi: 10.1016/0020-7519(73)90095-7. [DOI] [PubMed] [Google Scholar]
  13. Brun R., Jenni L., Schönenberger M., Schell K. F. In vitro cultivation of bloodstream forms of Trypanosoma brucei, T. rhodesiense, and T. gambiense. J Protozool. 1981 Nov;28(4):470–479. doi: 10.1111/j.1550-7408.1981.tb05322.x. [DOI] [PubMed] [Google Scholar]
  14. Brun R., Schönenberger Cultivation and in vitro cloning or procyclic culture forms of Trypanosoma brucei in a semi-defined medium. Short communication. Acta Trop. 1979 Sep;36(3):289–292. [PubMed] [Google Scholar]
  15. Brun R., Schönenberger M. Stimulating effect of citrate and cis-Aconitate on the transformation of Trypanosoma brucei bloodstream forms to procyclic forms in vitro. Z Parasitenkd. 1981;66(1):17–24. doi: 10.1007/BF00941941. [DOI] [PubMed] [Google Scholar]
  16. Cardoso de Almeida M. L., Turner M. J. The membrane form of variant surface glycoproteins of Trypanosoma brucei. Nature. 1983 Mar 24;302(5906):349–352. doi: 10.1038/302349a0. [DOI] [PubMed] [Google Scholar]
  17. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  18. Coudrier E., Reggio H., Louvard D. Immunolocalization of the 110,000 molecular weight cytoskeletal protein of intestinal microvilli. J Mol Biol. 1981 Oct 15;152(1):49–66. doi: 10.1016/0022-2836(81)90095-4. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  21. Evans D. A., Brown R. C. The utilization of glucose and proline by culture forms of Trypanosoma brucei. J Protozool. 1972 Nov;19(4):686–690. doi: 10.1111/j.1550-7408.1972.tb03561.x. [DOI] [PubMed] [Google Scholar]
  22. Evans D. A. Cyclical transmission of Trypanosoma brucei rhodesiense and Trypanosoma congolense by tsetse flies infected with culture-form procyclic trypanosomes. J Protozool. 1979 Aug;26(3):425–427. doi: 10.1111/j.1550-7408.1979.tb04648.x. [DOI] [PubMed] [Google Scholar]
  23. Ghiotto V., Brun R., Jenni L., Hecker H. Trypanosoma brucei: morphometric changes and loss of infectivity during transformation of bloodstream forms to procyclic culture forms in vitro. Exp Parasitol. 1979 Dec;48(3):447–456. doi: 10.1016/0014-4894(79)90129-2. [DOI] [PubMed] [Google Scholar]
  24. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Hoeijmakers J. H., Frasch A. C., Bernards A., Borst P., Cross G. A. Novel expression-linked copies of the genes for variant surface antigens in trypanosomes. Nature. 1980 Mar 6;284(5751):78–80. doi: 10.1038/284078a0. [DOI] [PubMed] [Google Scholar]
  27. Holder A. A. Carbohydrate is linked through ethanolamine to the C-terminal amino acid of Trypanosoma brucei variant surface glycoprotein. Biochem J. 1983 Jan 1;209(1):261–262. doi: 10.1042/bj2090261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Holder A. A., Cross G. A. Glycopeptides from variant surface glycoproteins of Trypanosoma Brucei. C-terminal location of antigenically cross-reacting carbohydrate moieties. Mol Biochem Parasitol. 1981 Feb;2(3-4):135–150. doi: 10.1016/0166-6851(81)90095-5. [DOI] [PubMed] [Google Scholar]
  29. Holder A. A., Freeman R. R. Biosynthesis and processing of a Plasmodium falciparum schizont antigen recognized by immune serum and a monoclonal antibody. J Exp Med. 1982 Nov 1;156(5):1528–1538. doi: 10.1084/jem.156.5.1528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Johnson J. G., Cross G. A. Selective cleavage of variant surface glycoproteins from Trypanosoma brucei. Biochem J. 1979 Mar 15;178(3):689–697. doi: 10.1042/bj1780689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  32. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  33. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pays E., Van Meirvenne N., Le Ray D., Steinert M. Gene duplication and transposition linked to antigenic variation in Trypanosoma brucei. Proc Natl Acad Sci U S A. 1981 May;78(5):2673–2677. doi: 10.1073/pnas.78.5.2673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. SEED J. R. ANTIGENIC SIMILARITY AMONG CULTURE FORMS OF THE 'BRUCEI' GROUP OF TRYPANOSOMES. Parasitology. 1964 Aug;54:593–596. doi: 10.1017/s0031182000082639. [DOI] [PubMed] [Google Scholar]
  37. Schöni R., Jenni L., Brun R. Cyclical transmission of in vitro cultivated bloodstream forms and procyclic trypomastigotes of trypanosoma brucei brucei by Glossina morsitans morsitans. Z Parasitenkd. 1982;68(1):1–5. doi: 10.1007/BF00926651. [DOI] [PubMed] [Google Scholar]
  38. Simpson L., Simpson A. M., Kidane G., Livingston L., Spithill T. W. The kinetoplast DNA of the hemoflagellate protozoa. Am J Trop Med Hyg. 1980 Sep;29(5 Suppl):1053–1063. doi: 10.4269/ajtmh.1980.29.1053. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Taylor D. W., Cross G. A. The synthesis of a variant-specific antigen by Trypanosoma brucei in vitro. Parasitology. 1977 Feb;74(1):47–60. doi: 10.1017/s0031182000047521. [DOI] [PubMed] [Google Scholar]
  41. Tetley L., Vickerman K., Moloo S. K. Absence of a surface coat from metacyclic Trypanosoma vivax: possible implications for vaccination against vivax trypanosomiasis. Trans R Soc Trop Med Hyg. 1981;75(3):409–414. doi: 10.1016/0035-9203(81)90106-1. [DOI] [PubMed] [Google Scholar]
  42. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Van der Ploeg L. H., Bernards A., Rijsewijk F. A., Borst P. Characterization of the DNA duplication-transposition that controls the expression of two genes for variant surface glycoproteins in Trypanosoma brucei. Nucleic Acids Res. 1982 Jan 22;10(2):593–609. doi: 10.1093/nar/10.2.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Van der Ploeg L. H., Liu A. Y., Michels P. A., De Lange T., Borst P., Majumder H. K., Weber H., Veeneman G. H., Van Boom J. RNA splicing is required to make the messenger RNA for a variant surface antigen in trypanosomes. Nucleic Acids Res. 1982 Jun 25;10(12):3591–3604. doi: 10.1093/nar/10.12.3591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Vickerman K., Luckins A. G. Localization of variable antigens in the surface coat of Trypanosoma brucei using ferritin conjugated antibody. Nature. 1969 Dec 13;224(5224):1125–1126. doi: 10.1038/2241125a0. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES