Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1995 Jun 15;14(12):2925–2934. doi: 10.1002/j.1460-2075.1995.tb07292.x

The 3'-terminal region of the mRNAs for VSG and procyclin can confer stage specificity to gene expression in Trypanosoma brucei.

M Berberof 1, L Vanhamme 1, P Tebabi 1, A Pays 1, D Jefferies 1, S Welburn 1, E Pays 1
PMCID: PMC398412  PMID: 7796818

Abstract

The variant surface glycoprotein (VSG) and procyclin are the respective major surface antigens of the bloodstream and the procyclic forms of Trypanosoma brucei. These proteins and their mRNAs are both the most abundant and absolutely characteristic of their respective life cycle stages. We show that the 3'-terminal region of these mRNAs regulates expression of a reporter gene in an inverse manner, depending on the developmental form of the parasite. In the case of VSG mRNA, the 97 nt sequence upstream from the polyadenylation site is responsible for these effects. The regulation occurs through a variation of mRNA abundance which is not due to a change in primary transcription. In the bloodstream form this effect is manifested by an increase in RNA stability, whereas in the procyclic form it seems to be related to a reduction in the efficiency of mRNA maturation. The 3'-end of VSG mRNA can obviate the 5- to 10-fold stimulation of transcription driven by the procyclin promoter during differentiation from the bloodstream to the procyclic form. The predominance of posttranscriptional over transcriptional controls is probably linked to the organization of the trypanosome genome in polycistronic transcription units.

Full text

PDF
2925

Images in this article

2929Image
on p.2929
2929Image
on p.2929
2930Image
on p.2930
2931Image
on p.2931
2932Image
on p.2932
2932Image
on p.2932

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. Aline R. F., Jr, Stuart K. Trypanosoma brucei: conserved sequence organization 3' to telomeric variant surface glycoprotein genes. Exp Parasitol. 1989 Jan;68(1):57–66. doi: 10.1016/0014-4894(89)90008-8. [DOI] [PubMed] [Google Scholar]
  3. Aly R., Argaman M., Halman S., Shapira M. A regulatory role for the 5' and 3' untranslated regions in differential expression of hsp83 in Leishmania. Nucleic Acids Res. 1994 Aug 11;22(15):2922–2929. doi: 10.1093/nar/22.15.2922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. 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]
  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. Clayton C. Developmental regulation of nuclear gene expression in Trypanosoma brucei. Prog Nucleic Acid Res Mol Biol. 1992;43:37–66. doi: 10.1016/s0079-6603(08)61043-0. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Coquelet H., Tebabi P., Pays A., Steinert M., Pays E. Trypanosoma brucei: enrichment by UV of intergenic transcripts from the variable surface glycoprotein gene expression site. Mol Cell Biol. 1989 Sep;9(9):4022–4025. doi: 10.1128/mcb.9.9.4022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Ehlers B., Czichos J., Overath P. RNA turnover in Trypanosoma brucei. Mol Cell Biol. 1987 Mar;7(3):1242–1249. doi: 10.1128/mcb.7.3.1242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Gibson W. C., Swinkels B. W., Borst P. Post-transcriptional control of the differential expression of phosphoglycerate kinase genes in Trypanosoma brucei. J Mol Biol. 1988 May 20;201(2):315–325. doi: 10.1016/0022-2836(88)90140-4. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Hehl A., Vassella E., Braun R., Roditi I. A conserved stem-loop structure in the 3' untranslated region of procyclin mRNAs regulates expression in Trypanosoma brucei. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):370–374. doi: 10.1073/pnas.91.1.370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Huang J., Van der Ploeg L. H. Requirement of a polypyrimidine tract for trans-splicing in trypanosomes: discriminating the PARP promoter from the immediately adjacent 3' splice acceptor site. EMBO J. 1991 Dec;10(12):3877–3885. doi: 10.1002/j.1460-2075.1991.tb04957.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hug M., Carruthers V. B., Hartmann C., Sherman D. S., Cross G. A., Clayton C. A possible role for the 3'-untranslated region in developmental regulation in Trypanosoma brucei. Mol Biochem Parasitol. 1993 Sep;61(1):87–95. doi: 10.1016/0166-6851(93)90161-p. [DOI] [PubMed] [Google Scholar]
  21. Hug M., Hotz H. R., Hartmann C., Clayton C. Hierarchies of RNA-processing signals in a trypanosome surface antigen mRNA precursor. Mol Cell Biol. 1994 Nov;14(11):7428–7435. doi: 10.1128/mcb.14.11.7428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Kapotas N., Bellofatto V. Differential response to RNA trans-splicing signals within the phosphoglycerate kinase gene cluster in Trypanosoma brucei. Nucleic Acids Res. 1993 Aug 25;21(17):4067–4072. doi: 10.1093/nar/21.17.4067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Koenig-Martin E., Yamage M., Roditi I. A procyclin-associated gene in Trypanosoma brucei encodes a polypeptide related to ESAG 6 and 7 proteins. Mol Biochem Parasitol. 1992 Oct;55(1-2):135–145. doi: 10.1016/0166-6851(92)90134-6. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. LeBowitz J. H., Smith H. Q., Rusche L., Beverley S. M. Coupling of poly(A) site selection and trans-splicing in Leishmania. Genes Dev. 1993 Jun;7(6):996–1007. doi: 10.1101/gad.7.6.996. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Matthews K. R., Tschudi C., Ullu E. A common pyrimidine-rich motif governs trans-splicing and polyadenylation of tubulin polycistronic pre-mRNA in trypanosomes. Genes Dev. 1994 Feb 15;8(4):491–501. doi: 10.1101/gad.8.4.491. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. 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]
  35. 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]
  36. 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]
  37. Pays E., Vanhamme L., Berberof M. Genetic controls for the expression of surface antigens in African trypanosomes. Annu Rev Microbiol. 1994;48:25–52. doi: 10.1146/annurev.mi.48.100194.000325. [DOI] [PubMed] [Google Scholar]
  38. Revelard P., Lips S., Pays E. Alternative splicing within and between alleles of the ATPase gene 1 locus of Trypanosoma brucei. Mol Biochem Parasitol. 1993 Nov;62(1):93–101. doi: 10.1016/0166-6851(93)90181-v. [DOI] [PubMed] [Google Scholar]
  39. Roditi I., Pearson T. W. The procyclin coat of African trypanosomes (or the not-so-naked trypanosome). Parasitol Today. 1990 Mar;6(3):79–82. doi: 10.1016/0169-4758(90)90216-q. [DOI] [PubMed] [Google Scholar]
  40. Roditi I., Schwarz H., Pearson T. W., Beecroft R. P., Liu M. K., Richardson J. P., Bühring H. J., Pleiss J., Bülow R., Williams R. O. Procyclin gene expression and loss of the variant surface glycoprotein during differentiation of Trypanosoma brucei. J Cell Biol. 1989 Feb;108(2):737–746. doi: 10.1083/jcb.108.2.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. 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]
  43. Schürch N., Hehl A., Vassella E., Braun R., Roditi I. Accurate polyadenylation of procyclin mRNAs in Trypanosoma brucei is determined by pyrimidine-rich elements in the intergenic regions. Mol Cell Biol. 1994 Jun;14(6):3668–3675. doi: 10.1128/mcb.14.6.3668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. 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]
  45. Vassella E., Braun R., Roditi I. Control of polyadenylation and alternative splicing of transcripts from adjacent genes in a procyclin expression site: a dual role for polypyrimidine tracts in trypanosomes? Nucleic Acids Res. 1994 Apr 25;22(8):1359–1364. doi: 10.1093/nar/22.8.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. 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]
  47. 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]
  48. 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 The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES