Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1993 Nov;13(11):7036–7044. doi: 10.1128/mcb.13.11.7036

Expression of a retroposon-like sequence upstream of the putative Trypanosoma brucei variant surface glycoprotein gene expression site promoter.

M J Lodes 1, B L Smiley 1, A W Stadnyk 1, J L Bennett 1, P J Myler 1, K Stuart 1
PMCID: PMC364765  PMID: 8413293

Abstract

We have cloned the region spanning the putative promoter from two variant surface glycoprotein gene expression sites that are at each end of chromosome M4 of Trypanosoma brucei IsTat 7. Both expression sites contain a retroposon-like sequence (ESR) pseudogene whose 3' end is approximately 30 bp upstream of the putative expression site promoter. The ESRs from both expression sites share considerable sequence homology and are related to LINE-like elements, especially the T. brucei ingi retroposon. Other ESRs are located on large, but not intermediate or mini-, chromosomes in the IsTaR 1 serodeme, and the total copy number is 10 to 20, similar to that estimated for variant surface glycoprotein expression sites. No DNA rearrangements in the vicinity of the ESR and putative expression site promoter were detected following antigenic switches in the IsTaR 1 serodeme. ESR transcripts are present in bloodstream, but not procyclic, forms. Variation in transcript size and sequence between bloodstream variant antigenic types implies that only the ESR from the active expression site is transcribed. This pattern of expression reflects that of sequences downstream of the putative expression site promoter, suggesting that the region of coordinately controlled expression extends upstream of this promoter.

Full text

PDF
7036

Images in this article

Selected References

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

  1. Aksoy S., Lalor T. M., Martin J., Van der Ploeg L. H., Richards F. F. Multiple copies of a retroposon interrupt spliced leader RNA genes in the African trypanosome, Trypanosoma gambiense. EMBO J. 1987 Dec 1;6(12):3819–3826. doi: 10.1002/j.1460-2075.1987.tb02718.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barry J. D., Turner C. M. The dynamics of antigenic variation and growth of African trypanosomes. Parasitol Today. 1991 Aug;7(8):207–211. doi: 10.1016/0169-4758(91)90143-c. [DOI] [PubMed] [Google Scholar]
  3. Cabot E. L., Beckenbach A. T. Simultaneous editing of multiple nucleic acid and protein sequences with ESEE. Comput Appl Biosci. 1989 Jul;5(3):233–234. doi: 10.1093/bioinformatics/5.3.233. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Cross G. A. Cellular and genetic aspects of antigenic variation in trypanosomes. Annu Rev Immunol. 1990;8:83–110. doi: 10.1146/annurev.iy.08.040190.000503. [DOI] [PubMed] [Google Scholar]
  6. Crozatier M., Van der Ploeg L. H., Johnson P. J., Gommers-Ampt J., Borst P. Structure of a telomeric expression site for variant specific surface antigens in Trypanosoma brucei. Mol Biochem Parasitol. 1990 Aug;42(1):1–12. doi: 10.1016/0166-6851(90)90107-w. [DOI] [PubMed] [Google Scholar]
  7. Cully D. F., Ip H. S., Cross G. A. Coordinate transcription of variant surface glycoprotein genes and an expression site associated gene family in Trypanosoma brucei. Cell. 1985 Aug;42(1):173–182. doi: 10.1016/s0092-8674(85)80113-6. [DOI] [PubMed] [Google Scholar]
  8. Donelson J. E., Rice-Ficht A. C. Molecular biology of trypanosome antigenic variation. Microbiol Rev. 1985 Jun;49(2):107–125. doi: 10.1128/mr.49.2.107-125.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gabriel A., Boeke J. D. Reverse transcriptase encoded by a retrotransposon from the trypanosomatid Crithidia fasciculata. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9794–9798. doi: 10.1073/pnas.88.21.9794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gottesdiener K. M., Goriparthi L., Masucci J. P., Van der Ploeg L. H. A proposed mechanism for promoter-associated DNA rearrangement events at a variant surface glycoprotein gene expression site. Mol Cell Biol. 1992 Oct;12(10):4784–4795. doi: 10.1128/mcb.12.10.4784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gottesdiener K., Chung H. M., Brown S. D., Lee M. G., Van der Ploeg L. H. Characterization of VSG gene expression site promoters and promoter-associated DNA rearrangement events. Mol Cell Biol. 1991 May;11(5):2467–2480. doi: 10.1128/mcb.11.5.2467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hattori M., Kuhara S., Takenaka O., Sakaki Y. L1 family of repetitive DNA sequences in primates may be derived from a sequence encoding a reverse transcriptase-related protein. Nature. 1986 Jun 5;321(6070):625–628. doi: 10.1038/321625a0. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Jefferies D., Tebabi P., Pays E. Transient activity assays of the Trypanosoma brucei variant surface glycoprotein gene promoter: control of gene expression at the posttranscriptional level. Mol Cell Biol. 1991 Jan;11(1):338–343. doi: 10.1128/mcb.11.1.338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kimmel B. E., ole-MoiYoi O. K., Young J. R. Ingi, a 5.2-kb dispersed sequence element from Trypanosoma brucei that carries half of a smaller mobile element at either end and has homology with mammalian LINEs. Mol Cell Biol. 1987 Apr;7(4):1465–1475. doi: 10.1128/mcb.7.4.1465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Marlor R. L., Parkhurst S. M., Corces V. G. The Drosophila melanogaster gypsy transposable element encodes putative gene products homologous to retroviral proteins. Mol Cell Biol. 1986 Apr;6(4):1129–1134. doi: 10.1128/mcb.6.4.1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Milhausen M., Nelson R. G., Parsons M., Newport G., Stuart K., Agabian N. Molecular characterization of initial variants from the IsTat I serodeme of Trypanosoma brucei. Mol Biochem Parasitol. 1983 Nov;9(3):241–254. doi: 10.1016/0166-6851(83)90100-7. [DOI] [PubMed] [Google Scholar]
  18. Mount S. M., Rubin G. M. Complete nucleotide sequence of the Drosophila transposable element copia: homology between copia and retroviral proteins. Mol Cell Biol. 1985 Jul;5(7):1630–1638. doi: 10.1128/mcb.5.7.1630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Murphy N. B., Pays A., Tebabi P., Coquelet H., Guyaux M., Steinert M., Pays E. Trypanosoma brucei repeated element with unusual structural and transcriptional properties. J Mol Biol. 1987 Jun 20;195(4):855–871. doi: 10.1016/0022-2836(87)90490-6. [DOI] [PubMed] [Google Scholar]
  20. Myler P. J., Aline R. F., Jr, Scholler J. K., Stuart K. D. Changes in telomere length associated with antigenic variation in Trypanosoma brucei. Mol Biochem Parasitol. 1988 Jun;29(2-3):243–250. doi: 10.1016/0166-6851(88)90079-5. [DOI] [PubMed] [Google Scholar]
  21. Myler P. J., Aline R. F., Jr, Scholler J. K., Stuart K. D. Multiple events associated with antigenic switching in Trypanosoma brucei. Mol Biochem Parasitol. 1988 Jun;29(2-3):227–241. doi: 10.1016/0166-6851(88)90078-3. [DOI] [PubMed] [Google Scholar]
  22. Pays E., Coquelet H., Tebabi P., Pays A., Jefferies D., Steinert M., Koenig E., Williams R. O., Roditi I. Trypanosoma brucei: constitutive activity of the VSG and procyclin gene promoters. EMBO J. 1990 Oct;9(10):3145–3151. doi: 10.1002/j.1460-2075.1990.tb07512.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pays E. Gene conversion in trypanosome antigenic variation. Prog Nucleic Acid Res Mol Biol. 1985;32:1–26. doi: 10.1016/s0079-6603(08)60344-x. [DOI] [PubMed] [Google Scholar]
  24. Pays E., Guyaux M., Aerts D., Van Meirvenne N., Steinert M. Telomeric reciprocal recombination as a possible mechanism for antigenic variation in trypanosomes. Nature. 1985 Aug 8;316(6028):562–564. doi: 10.1038/316562a0. [DOI] [PubMed] [Google Scholar]
  25. Pays E., Tebabi P., Pays A., Coquelet H., Revelard P., Salmon D., Steinert M. The genes and transcripts of an antigen gene expression site from T. brucei. Cell. 1989 Jun 2;57(5):835–845. doi: 10.1016/0092-8674(89)90798-8. [DOI] [PubMed] [Google Scholar]
  26. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Poch O., Sauvaget I., Delarue M., Tordo N. Identification of four conserved motifs among the RNA-dependent polymerase encoding elements. EMBO J. 1989 Dec 1;8(12):3867–3874. doi: 10.1002/j.1460-2075.1989.tb08565.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rogers J. H. The origin and evolution of retroposons. Int Rev Cytol. 1985;93:187–279. doi: 10.1016/s0074-7696(08)61375-3. [DOI] [PubMed] [Google Scholar]
  29. Scholler J. K., Aline R. F., Jr, Stuart K. D. Variant specific transcripts from the co-transposed segments of variant surface glycoprotein genes in Trypanosoma brucei. Mol Biochem Parasitol. 1988 May;29(1):89–103. doi: 10.1016/0166-6851(88)90123-5. [DOI] [PubMed] [Google Scholar]
  30. Scholler J. K., Reed S. G., Stuart K. Molecular karyotype of species and subspecies of Leishmania. Mol Biochem Parasitol. 1986 Sep;20(3):279–293. doi: 10.1016/0166-6851(86)90108-8. [DOI] [PubMed] [Google Scholar]
  31. Shea C., Glass D. J., Parangi S., Van der Ploeg L. H. Variant surface glycoprotein gene expression site switches in Trypanosoma brucei. J Biol Chem. 1986 May 5;261(13):6056–6063. [PubMed] [Google Scholar]
  32. Smiley B. L., Aline R. F., Jr, Myler P. J., Stuart K. A retroposon in the 5' flank of a Trypanosoma brucei VSG gene lacks insertional terminal repeats. Mol Biochem Parasitol. 1990 Aug;42(1):143–151. doi: 10.1016/0166-6851(90)90122-3. [DOI] [PubMed] [Google Scholar]
  33. Smiley B. L., Stadnyk A. W., Myler P. J., Stuart K. The trypanosome leucine repeat gene in the variant surface glycoprotein expression site encodes a putative metal-binding domain and a region resembling protein-binding domains of yeast, Drosophila, and mammalian proteins. Mol Cell Biol. 1990 Dec;10(12):6436–6444. doi: 10.1128/mcb.10.12.6436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Stuart K., Gobright E., Jenni L., Milhausen M., Thomashow L., Agabian N. The IsTaR 1 serodeme of Trypanosoma brucei: development of a new serodeme. J Parasitol. 1984 Oct;70(5):747–754. [PubMed] [Google Scholar]
  35. Thomashow L. S., Milhausen M., Rutter W. J., Agabian N. Tubulin genes are tandemly linked and clustered in the genome of trypanosoma brucei. Cell. 1983 Jan;32(1):35–43. doi: 10.1016/0092-8674(83)90494-4. [DOI] [PubMed] [Google Scholar]
  36. Van der Ploeg L. H. Control of antigenic variation in African trypanosomes. New Biol. 1991 Apr;3(4):324–330. [PubMed] [Google Scholar]
  37. Van der Werf A., Van Assel S., Aerts D., Steinert M., Pays E. Telomere interactions may condition the programming of antigen expression in Trypanosoma brucei. EMBO J. 1990 Apr;9(4):1035–1040. doi: 10.1002/j.1460-2075.1990.tb08207.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Villanueva M. S., Williams S. P., Beard C. B., Richards F. F., Aksoy S. A new member of a family of site-specific retrotransposons is present in the spliced leader RNA genes of Trypanosoma cruzi. Mol Cell Biol. 1991 Dec;11(12):6139–6148. doi: 10.1128/mcb.11.12.6139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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]
  40. Zomerdijk J. C., Kieft R., Shiels P. G., Borst P. Alpha-amanitin-resistant transcription units in trypanosomes: a comparison of promoter sequences for a VSG gene expression site and for the ribosomal RNA genes. Nucleic Acids Res. 1991 Oct 11;19(19):5153–5158. doi: 10.1093/nar/19.19.5153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zomerdijk J. C., Ouellette M., ten Asbroek A. L., Kieft R., Bommer A. M., Clayton C. E., Borst P. The promoter for a variant surface glycoprotein gene expression site in Trypanosoma brucei. EMBO J. 1990 Sep;9(9):2791–2801. doi: 10.1002/j.1460-2075.1990.tb07467.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES