Abstract
When the African trypanosome Trypanosoma brucei is taken up from mammals by a tse-tse fly, it replaces its variant surface glycoprotein (VSG) coat by a procyclin coat. Transcription of VSG genes stops in the fly, but transcription of sequences derived from the promoter area of the VSG expression site(s) remains high. Whether this is due to continuing high activity of one promoter or to low activity of many promoters was unclear. We have used the small differences between the sequences of different expression sites to show that multiple expression site promoters are active in insect form trypanosomes. This is confirmed by the low expression of single copy marker genes introduced into the transcribed area. However, if the expression site promoter is removed from the genomic location of the expression site and inserted in the non-transcribed spacer of the ribosomal DNA (rDNA), it is derepressed. Derepression of transcription can also be accomplished by replacing the promoter of an expression site by an rDNA promoter. We conclude that the down-regulation of VSG gene expression site promoters in insect form trypanosomes is affected by both the DNA sequence of the promoter and the genomic context in which it resides.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aparicio O. M., Billington B. L., Gottschling D. E. Modifiers of position effect are shared between telomeric and silent mating-type loci in S. cerevisiae. Cell. 1991 Sep 20;66(6):1279–1287. doi: 10.1016/0092-8674(91)90049-5. [DOI] [PubMed] [Google Scholar]
- Bernards A., De Lange T., Michels P. A., Liu A. Y., Huisman M. J., Borst P. Two modes of activation of a single surface antigen gene of Trypanosoma brucei. Cell. 1984 Jan;36(1):163–170. doi: 10.1016/0092-8674(84)90085-0. [DOI] [PubMed] [Google Scholar]
- 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]
- Bertrand G., Brunet P., Brun G., Pierre F., Thébault G. La prescription des anti-inflammatoires au cabinet. Rev Odontostomatol (Paris) 1979 Sep-Oct;8(5):289–292. [PubMed] [Google Scholar]
- Borst P. Discontinuous transcription and antigenic variation in trypanosomes. Annu Rev Biochem. 1986;55:701–732. doi: 10.1146/annurev.bi.55.070186.003413. [DOI] [PubMed] [Google Scholar]
- Chien C. T., Buck S., Sternglanz R., Shore D. Targeting of SIR1 protein establishes transcriptional silencing at HM loci and telomeres in yeast. Cell. 1993 Nov 5;75(3):531–541. doi: 10.1016/0092-8674(93)90387-6. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Cowell I. G. Repression versus activation in the control of gene transcription. Trends Biochem Sci. 1994 Jan;19(1):38–42. doi: 10.1016/0968-0004(94)90172-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Delauw M. F., Pays E., Steinert M., Aerts D., Van Meirvenne N., Le Ray D. Inactivation and reactivation of a variant-specific antigen gene in cyclically transmitted Trypanosoma brucei. EMBO J. 1985 Apr;4(4):989–993. doi: 10.1002/j.1460-2075.1985.tb03728.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dorn P. L., Aman R. A., Boothroyd J. C. Inhibition of protein synthesis results in super-induction of procyclin (PARP) RNA levels. Mol Biochem Parasitol. 1991 Jan;44(1):133–139. doi: 10.1016/0166-6851(91)90229-y. [DOI] [PubMed] [Google Scholar]
- Erondu N. E., Donelson J. E. Differential expression of two mRNAs from a single gene encoding an HMG1-like DNA binding protein of African trypanosomes. Mol Biochem Parasitol. 1992 Mar;51(1):111–118. doi: 10.1016/0166-6851(92)90206-y. [DOI] [PubMed] [Google Scholar]
- Gibson W., Garside L. Kinetoplast DNA minicircles are inherited from both parents in genetic hybrids of Trypanosoma brucei. Mol Biochem Parasitol. 1990 Aug;42(1):45–53. doi: 10.1016/0166-6851(90)90111-x. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- 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]
- Johnson P. J., Kooter J. M., Borst P. Inactivation of transcription by UV irradiation of T. brucei provides evidence for a multicistronic transcription unit including a VSG gene. Cell. 1987 Oct 23;51(2):273–281. doi: 10.1016/0092-8674(87)90154-1. [DOI] [PubMed] [Google Scholar]
- Kaster K. R., Burgett S. G., Rao R. N., Ingolia T. D. Analysis of a bacterial hygromycin B resistance gene by transcriptional and translational fusions and by DNA sequencing. Nucleic Acids Res. 1983 Oct 11;11(19):6895–6911. doi: 10.1093/nar/11.19.6895. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Krumm A., Meulia T., Groudine M. Common mechanisms for the control of eukaryotic transcriptional elongation. Bioessays. 1993 Oct;15(10):659–665. doi: 10.1002/bies.950151005. [DOI] [PubMed] [Google Scholar]
- Laird P. W., Kooter J. M., Loosbroek N., Borst P. Mature mRNAs of Trypanosoma brucei possess a 5' cap acquired by discontinuous RNA synthesis. Nucleic Acids Res. 1985 Jun 25;13(12):4253–4266. doi: 10.1093/nar/13.12.4253. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee M. G., Van der Ploeg L. H. Frequent independent duplicative transpositions activate a single VSG gene. Mol Cell Biol. 1987 Jan;7(1):357–364. doi: 10.1128/mcb.7.1.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lenardo M. J., Dorfman D. M., Reddy L. V., Donelson J. E. Characterization of the Trypanosoma brucei 5S ribosomal RNA gene and transcript: the 5S rRNA is a spliced-leader-independent species. Gene. 1985;35(1-2):131–141. doi: 10.1016/0378-1119(85)90165-9. [DOI] [PubMed] [Google Scholar]
- Ligtenberg M. J., Bitter W., Kieft R., Steverding D., Janssen H., Calafat J., Borst P. Reconstitution of a surface transferrin binding complex in insect form Trypanosoma brucei. EMBO J. 1994 Jun 1;13(11):2565–2573. doi: 10.1002/j.1460-2075.1994.tb06546.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Meulia T., Krumm A., Groudine M. Distinct properties of c-myc transcriptional elongation are revealed in Xenopus oocytes and mammalian cells and by template titration, 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), and promoter mutagenesis. Mol Cell Biol. 1993 Sep;13(9):5647–5658. doi: 10.1128/mcb.13.9.5647. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Pays E., Steinert M. Control of antigen gene expression in African trypanosomes. Annu Rev Genet. 1988;22:107–126. doi: 10.1146/annurev.ge.22.120188.000543. [DOI] [PubMed] [Google Scholar]
- 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]
- Renauld H., Aparicio O. M., Zierath P. D., Billington B. L., Chhablani S. K., Gottschling D. E. Silent domains are assembled continuously from the telomere and are defined by promoter distance and strength, and by SIR3 dosage. Genes Dev. 1993 Jul;7(7A):1133–1145. doi: 10.1101/gad.7.7a.1133. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Rougvie A. E., Lis J. T. The RNA polymerase II molecule at the 5' end of the uninduced hsp70 gene of D. melanogaster is transcriptionally engaged. Cell. 1988 Sep 9;54(6):795–804. doi: 10.1016/s0092-8674(88)91087-2. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- 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]
- Van der Ploeg L. H. Control of antigenic variation in African trypanosomes. New Biol. 1991 Apr;3(4):324–330. [PubMed] [Google Scholar]
- Van der Ploeg L. H., Smith C. L., Polvere R. I., Gottesdiener K. M. Improved separation of chromosome-sized DNA from Trypanosoma brucei, stock 427-60. Nucleic Acids Res. 1989 Apr 25;17(8):3217–3227. doi: 10.1093/nar/17.8.3217. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van der Ploeg L. H., Valerio D., De Lange T., Bernards A., Borst P., Grosveld F. G. An analysis of cosmid clones of nuclear DNA from Trypanosoma brucei shows that the genes for variant surface glycoproteins are clustered in the genome. Nucleic Acids Res. 1982 Oct 11;10(19):5905–5923. doi: 10.1093/nar/10.19.5905. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vickerman K. Antigenic variation in trypanosomes. Nature. 1978 Jun 22;273(5664):613–617. doi: 10.1038/273613a0. [DOI] [PubMed] [Google Scholar]
- Vickerman K. Developmental cycles and biology of pathogenic trypanosomes. Br Med Bull. 1985 Apr;41(2):105–114. doi: 10.1093/oxfordjournals.bmb.a072036. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- 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]
- ten Asbroek A. L., Mol C. A., Kieft R., Borst P. Stable transformation of Trypanosoma brucei. Mol Biochem Parasitol. 1993 May;59(1):133–142. doi: 10.1016/0166-6851(93)90014-o. [DOI] [PubMed] [Google Scholar]
- 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]