Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1995 Mar 25;23(6):1010–1018. doi: 10.1093/nar/23.6.1010

PARP promoter-mediated activation of a VSG expression site promoter in insect form Trypanosoma brucei.

T P Urményi 1, L H Van der Ploeg 1
PMCID: PMC306799  PMID: 7731788

Abstract

In trypanosomes the rRNA, PARP and VSG gene promoters mediate alpha-amanitin-resistant transcription of protein coding genes, presumably by RNA polymerase (pol) I. We compared the activity of PARP and VSG promoters integrated at one of the alleles of the largest subunit of pol II genes in insect form trypanosomes. Even though both promoters are roughly equally active in transient transformation assays in insect form trypanosomes, only the PARP promoter functioned effectively when integrated at the pol II largest subunit or other loci. Promoter activity in transient transformation assays is therefore not necessarily predictive of transcriptional activity once integrated into the trypanosome genome. The integrated fully active PARP promoter could upregulate in cis an otherwise poorly active integrated VSG promoter. The PARP promoter nucleotide sequence elements responsible for VSG promoter activation coincided with most of the important PARP promoter elements mapped previously by linker scanning mutagenesis, indicating that it is not a single unique promoter element that was responsible for VSG promoter activation. The data suggest that PARP promoter-mediated activation of the VSG promoter does not result from complementation of the VSG promoter with a single insect form-specific transcription factor whose binding site is missing from the VSG promoter and present in the PARP promoter. We favor a model in which chromatin structure at the locus is altered by the PARP promoter, allowing VSG promoter activation in insect form trypanosomes. We discuss the significance of these observations for the control of VSG promoters in insect form trypanosomes.

Full text

PDF
1010

Images in this article

1014Image
on p.1014
1015Image
on p.1015

Selected References

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

  1. Alarcon C. M., Son H. J., Hall T., Donelson J. E. A monocistronic transcript for a trypanosome variant surface glycoprotein. Mol Cell Biol. 1994 Aug;14(8):5579–5591. doi: 10.1128/mcb.14.8.5579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Borst P. Molecular genetics of antigenic variation. Immunol Today. 1991 Mar;12(3):A29–A33. doi: 10.1016/S0167-5699(05)80009-X. [DOI] [PubMed] [Google Scholar]
  4. Brown S. D., Huang J., Van der Ploeg L. H. The promoter for the procyclic acidic repetitive protein (PARP) genes of Trypanosoma brucei shares features with RNA polymerase I promoters. Mol Cell Biol. 1992 Jun;12(6):2644–2652. doi: 10.1128/mcb.12.6.2644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Cereghini S., Yaniv M. Assembly of transfected DNA into chromatin: structural changes in the origin-promoter-enhancer region upon replication. EMBO J. 1984 Jun;3(6):1243–1253. doi: 10.1002/j.1460-2075.1984.tb01959.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chung H. M., Lee M. G., Dietrich P., Huang J., Van der Ploeg L. H. Disruption of largest subunit RNA polymerase II genes in Trypanosoma brucei. Mol Cell Biol. 1993 Jun;13(6):3734–3743. doi: 10.1128/mcb.13.6.3734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. 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]
  11. Elledge S. J., Davis R. W. Position and density effects on repression by stationary and mobile DNA-binding proteins. Genes Dev. 1989 Feb;3(2):185–197. doi: 10.1101/gad.3.2.185. [DOI] [PubMed] [Google Scholar]
  12. Evers R., Hammer A., Köck J., Jess W., Borst P., Mémet S., Cornelissen A. W. Trypanosoma brucei contains two RNA polymerase II largest subunit genes with an altered C-terminal domain. Cell. 1989 Feb 24;56(4):585–597. doi: 10.1016/0092-8674(89)90581-3. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Gottesdiener K., Garciá-Anoveros J., Lee M. G., Van der Ploeg L. H. Chromosome organization of the protozoan Trypanosoma brucei. Mol Cell Biol. 1990 Nov;10(11):6079–6083. doi: 10.1128/mcb.10.11.6079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Ingelbrecht I., Breyne P., Vancompernolle K., Jacobs A., Van Montagu M., Depicker A. Transcriptional interference in transgenic plants. Gene. 1991 Dec 30;109(2):239–242. doi: 10.1016/0378-1119(91)90614-h. [DOI] [PubMed] [Google Scholar]
  18. Janz L., Clayton C. The PARP and rRNA promoters of Trypanosoma brucei are composed of dissimilar sequence elements that are functionally interchangeable. Mol Cell Biol. 1994 Sep;14(9):5804–5811. doi: 10.1128/mcb.14.9.5804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. 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]
  22. Kooter J. M., Borst P. Alpha-amanitin-insensitive transcription of variant surface glycoprotein genes provides further evidence for discontinuous transcription in trypanosomes. Nucleic Acids Res. 1984 Dec 21;12(24):9457–9472. doi: 10.1093/nar/12.24.9457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. 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]
  26. Lee M. G., van der Ploeg L. H. The hygromycin B-resistance-encoding gene as a selectable marker for stable transformation of Trypanosoma brucei. Gene. 1991 Sep 15;105(2):255–257. doi: 10.1016/0378-1119(91)90159-9. [DOI] [PubMed] [Google Scholar]
  27. Minchiotti G., Di Nocera P. P. Convergent transcription initiates from oppositely oriented promoters within the 5' end regions of Drosophila melanogaster F elements. Mol Cell Biol. 1991 Oct;11(10):5171–5180. doi: 10.1128/mcb.11.10.5171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mowatt M. R., Clayton C. E. Developmental regulation of a novel repetitive protein of Trypanosoma brucei. Mol Cell Biol. 1987 Aug;7(8):2838–2844. doi: 10.1128/mcb.7.8.2838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mowatt M. R., Clayton C. E. Polymorphism in the procyclic acidic repetitive protein gene family of Trypanosoma brucei. Mol Cell Biol. 1988 Oct;8(10):4055–4062. doi: 10.1128/mcb.8.10.4055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mowatt M. R., Wisdom G. S., Clayton C. E. Variation of tandem repeats in the developmentally regulated procyclic acidic repetitive proteins of Trypanosoma brucei. Mol Cell Biol. 1989 Mar;9(3):1332–1335. doi: 10.1128/mcb.9.3.1332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nellen W., Lichtenstein C. What makes an mRNA anti-sense-itive? Trends Biochem Sci. 1993 Nov;18(11):419–423. doi: 10.1016/0968-0004(93)90137-c. [DOI] [PubMed] [Google Scholar]
  32. Paranjape S. M., Kamakaka R. T., Kadonaga J. T. Role of chromatin structure in the regulation of transcription by RNA polymerase II. Annu Rev Biochem. 1994;63:265–297. doi: 10.1146/annurev.bi.63.070194.001405. [DOI] [PubMed] [Google Scholar]
  33. Patnaik P. K., Kulkarni S. K., Cross G. A. Autonomously replicating single-copy episomes in Trypanosoma brucei show unusual stability. EMBO J. 1993 Jun;12(6):2529–2538. doi: 10.1002/j.1460-2075.1993.tb05908.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. 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]
  36. Pays E. Genetics of antigenic variation in African trypanosomes. Res Microbiol. 1991 Jul-Aug;142(6):731–735. doi: 10.1016/0923-2508(91)90088-r. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Pribyl T. M., Martinson H. G. Transcription termination at the chicken beta H-globin gene. Mol Cell Biol. 1988 Dec;8(12):5369–5377. doi: 10.1128/mcb.8.12.5369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Reeves R., Gorman C. M., Howard B. Minichromosome assembly of non-integrated plasmid DNA transfected into mammalian cells. Nucleic Acids Res. 1985 May 24;13(10):3599–3615. doi: 10.1093/nar/13.10.3599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Richardson J. P., Beecroft R. P., Tolson D. L., Liu M. K., Pearson T. W. Procyclin: an unusual immunodominant glycoprotein surface antigen from the procyclic stage of African trypanosomes. Mol Biochem Parasitol. 1988 Dec;31(3):203–216. doi: 10.1016/0166-6851(88)90150-8. [DOI] [PubMed] [Google Scholar]
  41. Roditi I., Carrington M., Turner M. Expression of a polypeptide containing a dipeptide repeat is confined to the insect stage of Trypanosoma brucei. Nature. 1987 Jan 15;325(6101):272–274. doi: 10.1038/325272a0. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. 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]
  44. Rudenko G., Bishop D., Gottesdiener K., Van der Ploeg L. H. Alpha-amanitin resistant transcription of protein coding genes in insect and bloodstream form Trypanosoma brucei. EMBO J. 1989 Dec 20;8(13):4259–4263. doi: 10.1002/j.1460-2075.1989.tb08611.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. 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]
  46. Rudenko G., Chung H. M., Pham V. P., Van der Ploeg L. H. RNA polymerase I can mediate expression of CAT and neo protein-coding genes in Trypanosoma brucei. EMBO J. 1991 Nov;10(11):3387–3397. doi: 10.1002/j.1460-2075.1991.tb04903.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. 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]
  48. Rudenko G., Lee M. G., Van der Ploeg L. H. The PARP and VSG genes of Trypanosoma brucei do not resemble RNA polymerase II transcription units in sensitivity to Sarkosyl in nuclear run-on assays. Nucleic Acids Res. 1992 Jan 25;20(2):303–306. doi: 10.1093/nar/20.2.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Rudenko G., Van der Ploeg L. H. Transcription of telomere repeats in protozoa. EMBO J. 1989 Sep;8(9):2633–2638. doi: 10.1002/j.1460-2075.1989.tb08403.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Saha S., Haggård-Ljungquist E., Nordström K. The cox protein of bacteriophage P2 inhibits the formation of the repressor protein and autoregulates the early operon. EMBO J. 1987 Oct;6(10):3191–3199. doi: 10.1002/j.1460-2075.1987.tb02631.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Scheer U., Benavente R. Functional and dynamic aspects of the mammalian nucleolus. Bioessays. 1990 Jan;12(1):14–21. doi: 10.1002/bies.950120104. [DOI] [PubMed] [Google Scholar]
  52. Shea C., Lee M. G., Van der Ploeg L. H. VSG gene 118 is transcribed from a cotransposed pol I-like promoter. Cell. 1987 Aug 14;50(4):603–612. doi: 10.1016/0092-8674(87)90033-x. [DOI] [PubMed] [Google Scholar]
  53. Sherman D. R., Janz L., Hug M., Clayton C. Anatomy of the parp gene promoter of Trypanosoma brucei. EMBO J. 1991 Nov;10(11):3379–3386. doi: 10.1002/j.1460-2075.1991.tb04902.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Smith J. L., Levin J. R., Ingles C. J., Agabian N. In trypanosomes the homolog of the largest subunit of RNA polymerase II is encoded by two genes and has a highly unusual C-terminal domain structure. Cell. 1989 Mar 10;56(5):815–827. doi: 10.1016/0092-8674(89)90686-7. [DOI] [PubMed] [Google Scholar]
  55. Sollner-Webb B., Mougey E. B. News from the nucleolus: rRNA gene expression. Trends Biochem Sci. 1991 Feb;16(2):58–62. doi: 10.1016/0968-0004(91)90025-q. [DOI] [PubMed] [Google Scholar]
  56. 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]
  57. Van der Ploeg L. H. Control of antigenic variation in African trypanosomes. New Biol. 1991 Apr;3(4):324–330. [PubMed] [Google Scholar]
  58. Van der Ploeg L. H., Gottesdiener K., Lee M. G. Antigenic variation in African trypanosomes. Trends Genet. 1992 Dec;8(12):452–457. doi: 10.1016/0168-9525(92)90330-7. [DOI] [PubMed] [Google Scholar]
  59. 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]
  60. White T. C., Rudenko G., Borst P. Three small RNAs within the 10 kb trypanosome rRNA transcription unit are analogous to domain VII of other eukaryotic 28S rRNAs. Nucleic Acids Res. 1986 Dec 9;14(23):9471–9489. doi: 10.1093/nar/14.23.9471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Williams T., Fried M. A mouse locus at which transcription from both DNA strands produces mRNAs complementary at their 3' ends. Nature. 1986 Jul 17;322(6076):275–279. doi: 10.1038/322275a0. [DOI] [PubMed] [Google Scholar]
  62. Zaret K. S., Sherman F. DNA sequence required for efficient transcription termination in yeast. Cell. 1982 Mar;28(3):563–573. doi: 10.1016/0092-8674(82)90211-2. [DOI] [PubMed] [Google Scholar]
  63. Zomerdijk J. C., Kieft R., Borst P. Efficient production of functional mRNA mediated by RNA polymerase I in Trypanosoma brucei. Nature. 1991 Oct 24;353(6346):772–775. doi: 10.1038/353772a0. [DOI] [PubMed] [Google Scholar]
  64. Zomerdijk J. C., Kieft R., Borst P. Insertion of the promoter for a variant surface glycoprotein gene expression site in an RNA polymerase II transcription unit of procyclic Trypanosoma brucei. Mol Biochem Parasitol. 1993 Feb;57(2):295–304. doi: 10.1016/0166-6851(93)90205-c. [DOI] [PubMed] [Google Scholar]
  65. 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]
  66. 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]
  67. 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 Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES