Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1995 Jun 11;23(11):1862–1869. doi: 10.1093/nar/23.11.1862

Stimuli of differentiation regulate RNA elongation in the transcription units for the major stage-specific antigens of Trypanosoma brucei.

L Vanhamme 1, M Berberof 1, D Le Ray 1, E Pays 1
PMCID: PMC306955  PMID: 7596810

Abstract

In Trypanosoma brucei, the mutually exclusive expression of the major surface antigens, the variant surface glycoprotein (VSG) of the bloodstream form and procyclin of the procyclic form, is due to a stage-specific accumulation of the respective mRNAs. Through the targeting of a reporter construct in the procyclin promoter region, we show that independently of any selection pressure, a relatively high level of transcription (approximately 10%) occurs from the procyclin promoter in the bloodstream form. This transcription leads to the production of detectable amounts of polyadenylated mRNAs. However, RNA elongation in the procyclin transcription unit is down-regulated at this stage. Transcription elongation in the procyclin and VSG units is inversely controlled by the combination of factors which cause the differentiation of bloodstream into procyclic forms in vitro. These factors include temperature, citrate/cis-aconitate and the incubation medium. Our results suggest that inverse regulations of primary transcription in the VSG and procyclin units are early events that underly the differentiation of the parasite.

Full text

PDF
1862

Images in this article

1864Image
on p.1864
1864Image
on p.1864
1865Image
on p.1865
1865Image
on p.1865
1867Image
on p.1867

Selected References

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

  1. Alexandre S., Guyaux M., Murphy N. B., Coquelet H., Pays A., Steinert M., Pays E. Putative genes of a variant-specific antigen gene transcription unit in Trypanosoma brucei. Mol Cell Biol. 1988 Jun;8(6):2367–2378. doi: 10.1128/mcb.8.6.2367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baltz T., Baltz D., Giroud C., Crockett J. Cultivation in a semi-defined medium of animal infective forms of Trypanosoma brucei, T. equiperdum, T. evansi, T. rhodesiense and T. gambiense. EMBO J. 1985 May;4(5):1273–1277. doi: 10.1002/j.1460-2075.1985.tb03772.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bellofatto V., Cross G. A. Expression of a bacterial gene in a trypanosomatid protozoan. Science. 1989 Jun 9;244(4909):1167–1169. doi: 10.1126/science.2499047. [DOI] [PubMed] [Google Scholar]
  4. Ben Amar M. F., Pays A., Tebabi P., Dero B., Seebeck T., Steinert M., Pays E. Structure and transcription of the actin gene of Trypanosoma brucei. Mol Cell Biol. 1988 May;8(5):2166–2176. doi: 10.1128/mcb.8.5.2166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berberof M., Pays A., Pays E. A similar gene is shared by both the variant surface glycoprotein and procyclin gene transcription units of Trypanosoma brucei. Mol Cell Biol. 1991 Mar;11(3):1473–1479. doi: 10.1128/mcb.11.3.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Berberof M., Vanhamme L., Tebabi P., Pays A., Jefferies D., Welburn S., Pays E. The 3'-terminal region of the mRNAs for VSG and procyclin can confer stage specificity to gene expression in Trypanosoma brucei. EMBO J. 1995 Jun 15;14(12):2925–2934. doi: 10.1002/j.1460-2075.1995.tb07292.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Chung H. M., Lee M. G., Van der Ploeg L. H. RNA polymerase I-mediated protein-coding gene expression in Trypanosoma brucei. Parasitol Today. 1992 Dec;8(12):414–418. doi: 10.1016/0169-4758(92)90194-7. [DOI] [PubMed] [Google Scholar]
  9. Clayton C. E., Fueri J. P., Itzhaki J. E., Bellofatto V., Sherman D. R., Wisdom G. S., Vijayasarathy S., Mowatt M. R. Transcription of the procyclic acidic repetitive protein genes of Trypanosoma brucei. Mol Cell Biol. 1990 Jun;10(6):3036–3047. doi: 10.1128/mcb.10.6.3036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Coquelet H., Steinert M., Pays E. Ultraviolet irradiation inhibits RNA decay and modifies ribosomal RNA processing in Trypanosoma brucei. Mol Biochem Parasitol. 1991 Jan;44(1):33–42. doi: 10.1016/0166-6851(91)90218-u. [DOI] [PubMed] [Google Scholar]
  11. Czichos J., Nonnengaesser C., Overath P. Trypanosoma brucei: cis-aconitate and temperature reduction as triggers of synchronous transformation of bloodstream to procyclic trypomastigotes in vitro. Exp Parasitol. 1986 Oct;62(2):283–291. doi: 10.1016/0014-4894(86)90033-0. [DOI] [PubMed] [Google Scholar]
  12. Eid J., Sollner-Webb B. Stable integrative transformation of Trypanosoma brucei that occurs exclusively by homologous recombination. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2118–2121. doi: 10.1073/pnas.88.6.2118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jefferies D., Tebabi P., Le Ray D., Pays E. The ble resistance gene as a new selectable marker for Trypanosoma brucei: fly transmission of stable procyclic transformants to produce antibiotic resistant bloodstream forms. Nucleic Acids Res. 1993 Jan 25;21(2):191–195. doi: 10.1093/nar/21.2.191. [DOI] [PMC free article] [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. Kooter J. M., van der Spek H. J., Wagter R., d'Oliveira C. E., van der Hoeven F., Johnson P. J., Borst P. The anatomy and transcription of a telomeric expression site for variant-specific surface antigens in T. brucei. Cell. 1987 Oct 23;51(2):261–272. doi: 10.1016/0092-8674(87)90153-x. [DOI] [PubMed] [Google Scholar]
  16. König E., Delius H., Carrington M., Williams R. O., Roditi I. Duplication and transcription of procyclin genes in Trypanosoma brucei. Nucleic Acids Res. 1989 Nov 11;17(21):8727–8739. doi: 10.1093/nar/17.21.8727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lee M. G., Van der Ploeg L. H. Homologous recombination and stable transfection in the parasitic protozoan Trypanosoma brucei. Science. 1990 Dec 14;250(4987):1583–1587. doi: 10.1126/science.2177225. [DOI] [PubMed] [Google Scholar]
  18. Lips S., Revelard P., Pays E. Identification of a new expression site-associated gene in the complete 30.5 kb sequence from the AnTat 1.3A variant surface protein gene expression site of Trypanosoma brucei. Mol Biochem Parasitol. 1993 Nov;62(1):135–137. doi: 10.1016/0166-6851(93)90189-5. [DOI] [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. Pays E., Coquelet H., Pays A., Tebabi P., Steinert M. Trypanosoma brucei: posttranscriptional control of the variable surface glycoprotein gene expression site. Mol Cell Biol. 1989 Sep;9(9):4018–4021. doi: 10.1128/mcb.9.9.4018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Pays E., Hanocq-Quertier J., Hanocq F., Van Assel S., Nolan D., Rolin S. Abrupt RNA changes precede the first cell division during the differentiation of Trypanosoma brucei bloodstream forms into procyclic forms in vitro. Mol Biochem Parasitol. 1993 Sep;61(1):107–114. doi: 10.1016/0166-6851(93)90163-r. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Rudenko G., Blundell P. A., Taylor M. C., Kieft R., Borst P. VSG gene expression site control in insect form Trypanosoma brucei. EMBO J. 1994 Nov 15;13(22):5470–5482. doi: 10.1002/j.1460-2075.1994.tb06882.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rudenko G., Le Blancq S., Smith J., Lee M. G., Rattray A., Van der Ploeg L. H. Procyclic acidic repetitive protein (PARP) genes located in an unusually small alpha-amanitin-resistant transcription unit: PARP promoter activity assayed by transient DNA transfection of Trypanosoma brucei. Mol Cell Biol. 1990 Jul;10(7):3492–3504. doi: 10.1128/mcb.10.7.3492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sommer J. M., Cheng Q. L., Keller G. A., Wang C. C. In vivo import of firefly luciferase into the glycosomes of Trypanosoma brucei and mutational analysis of the C-terminal targeting signal. Mol Biol Cell. 1992 Jul;3(7):749–759. doi: 10.1091/mbc.3.7.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ziegelbauer K., Quinten M., Schwarz H., Pearson T. W., Overath P. Synchronous differentiation of Trypanosoma brucei from bloodstream to procyclic forms in vitro. Eur J Biochem. 1990 Sep 11;192(2):373–378. doi: 10.1111/j.1432-1033.1990.tb19237.x. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. 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]
  30. ten Asbroek A. L., Ouellette M., Borst P. Targeted insertion of the neomycin phosphotransferase gene into the tubulin gene cluster of Trypanosoma brucei. Nature. 1990 Nov 8;348(6297):174–175. doi: 10.1038/348174a0. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES