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. 1985 Mar;5(3):545–553. doi: 10.1128/mcb.5.3.545

Structure and transcription of a telomeric surface antigen gene of Trypanosoma brucei.

A Bernards, J M Kooter, P Borst
PMCID: PMC366747  PMID: 3879972

Abstract

The gene encoding variant surface glycoprotein 221 in Trypanosoma brucei is located adjacent to a chromosome end and can be activated with or without a concomitant gene duplication. To test whether transcription initiates within the cloned segment of the 221 gene, we analyzed nascent and stable transcripts. We show here that the 221 coding region and 8.5 kilobases of adjacent upstream DNA are transcribed into nascent RNA at a similar rate when gene 221 is activated without duplication. Since only part of this transcribed upstream segment is transferred with the coding region to another telomere upon duplicative activation of gene 221, we infer that initiation of variant surface glycoprotein gene transcription occurs outside the gene segment that moves into an expression site by gene conversion. Our analysis shows that part of the variant surface glycoprotein 221 transcription unit consists of an unusual 3.5-kilobase tandem array of ca. 50 repeat segments and that a rearrangement in this array accompanies the nonduplicative activation of gene 221. A variant surface glycoprotein pseudogene is located within the transcription unit of gene 221, and we discuss models that account for this unusual situation.

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Selected References

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  1. Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bernards A., Michels P. A., Lincke C. R., Borst P. Growth of chromosome ends in multiplying trypanosomes. Nature. 1983 Jun 16;303(5918):592–597. doi: 10.1038/303592a0. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Bernards A., van Harten-Loosbroek N., Borst P. Modification of telomeric DNA in Trypanosoma brucei; a role in antigenic variation? Nucleic Acids Res. 1984 May 25;12(10):4153–4170. doi: 10.1093/nar/12.10.4153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Blackburn E. H., Challoner P. B. Identification of a telomeric DNA sequence in Trypanosoma brucei. Cell. 1984 Feb;36(2):447–457. doi: 10.1016/0092-8674(84)90238-1. [DOI] [PubMed] [Google Scholar]
  8. Boothroyd J. C., Cross G. A. Transcripts coding for variant surface glycoproteins of Trypanosoma brucei have a short, identical exon at their 5' end. Gene. 1982 Dec;20(2):281–289. doi: 10.1016/0378-1119(82)90046-4. [DOI] [PubMed] [Google Scholar]
  9. Boothroyd J. C., Paynter C. A., Coleman S. L., Cross G. A. Complete nucleotide sequence of complementary DNA coding for a variant surface glycoprotein from Trypanosoma brucei. J Mol Biol. 1982 May 25;157(3):547–556. doi: 10.1016/0022-2836(82)90475-2. [DOI] [PubMed] [Google Scholar]
  10. Borst P., Bernards A., van der Ploeg L. H., Michels P. A., Liu A. Y., de Lange T., Kooter J. M. The control of variant surface antigen synthesis in trypanosomes. Eur J Biochem. 1983 Dec 15;137(3):383–389. doi: 10.1111/j.1432-1033.1983.tb07840.x. [DOI] [PubMed] [Google Scholar]
  11. Campbell D. A., Thornton D. A., Boothroyd J. C. Apparent discontinuous transcription of Trypanosoma brucei variant surface antigen genes. 1984 Sep 27-Oct 3Nature. 311(5984):350–355. doi: 10.1038/311350a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Campbell D. A., van Bree M. P., Boothroyd J. C. The 5'-limit of transposition and upstream barren region of a trypanosome VSG gene: tandem 76 base-pair repeats flanking (TAA)90. Nucleic Acids Res. 1984 Mar 26;12(6):2759–2774. doi: 10.1093/nar/12.6.2759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cross G. A. Identification, purification and properties of clone-specific glycoprotein antigens constituting the surface coat of Trypanosoma brucei. Parasitology. 1975 Dec;71(3):393–417. doi: 10.1017/s003118200004717x. [DOI] [PubMed] [Google Scholar]
  14. De Lange T., Berkvens T. M., Veerman H. J., Frasch A. C., Barry J. D., Borst P. Comparison of the genes coding for the common 5' terminal sequence of messenger RNAs in three trypanosome species. Nucleic Acids Res. 1984 Jun 11;12(11):4431–4443. doi: 10.1093/nar/12.11.4431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. De Lange T., Borst P. Genomic environment of the expression-linked extra copies of genes for surface antigens of Trypanosoma brucei resembles the end of a chromosome. Nature. 1982 Sep 30;299(5882):451–453. doi: 10.1038/299451a0. [DOI] [PubMed] [Google Scholar]
  16. De Lange T., Kooter J. M., Michels P. A., Borst P. Telomere conversion in trypanosomes. Nucleic Acids Res. 1983 Dec 10;11(23):8149–8165. doi: 10.1093/nar/11.23.8149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. De Lange T., Liu A. Y., Van der Ploeg L. H., Borst P., Tromp M. C., Van Boom J. H. Tandem repetition of the 5' mini-exon of variant surface glycoprotein genes: a multiple promoter for VSG gene transcription? Cell. 1983 Oct;34(3):891–900. doi: 10.1016/0092-8674(83)90546-9. [DOI] [PubMed] [Google Scholar]
  18. De Lange T., Michels P. A., Veerman H. J., Cornelissen A. W., Borst P. Many trypanosome messenger RNAs share a common 5' terminal sequence. Nucleic Acids Res. 1984 May 11;12(9):3777–3790. doi: 10.1093/nar/12.9.3777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Frasch A. C., Borst P., Van den Burg J. Rapid evolution of genes coding for variant surface glycoproteins in trypanosomes. Gene. 1982 Feb;17(2):197–211. doi: 10.1016/0378-1119(82)90073-7. [DOI] [PubMed] [Google Scholar]
  20. Groudine M., Peretz M., Weintraub H. Transcriptional regulation of hemoglobin switching in chicken embryos. Mol Cell Biol. 1981 Mar;1(3):281–288. doi: 10.1128/mcb.1.3.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Kooter J. M., De Lange T., Borst P. Discontinuous synthesis of mRNA in trypanosomes. EMBO J. 1984 Oct;3(10):2387–2392. doi: 10.1002/j.1460-2075.1984.tb02144.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lanham S. M. Separation of trypanosomes from the blood of infected rats and mice by anion-exchangers. Nature. 1968 Jun 29;218(5148):1273–1274. doi: 10.1038/2181273a0. [DOI] [PubMed] [Google Scholar]
  24. Laurent M., Pays E., Magnus E., Van Meirvenne N., Matthyssens G., Williams R. O., Steinert M. DNA rearrangements linked to expression of a predominant surface antigen gene of trypanosomes. Nature. 1983 Mar 17;302(5905):263–266. doi: 10.1038/302263a0. [DOI] [PubMed] [Google Scholar]
  25. Liu A. Y., Van der Ploeg L. H., Rijsewijk F. A., Borst P. The transposition unit of variant surface glycoprotein gene 118 of Trypanosoma brucei. Presence of repeated elements at its border and absence of promoter-associated sequences. J Mol Biol. 1983 Jun 15;167(1):57–75. doi: 10.1016/s0022-2836(83)80034-5. [DOI] [PubMed] [Google Scholar]
  26. Longacre S., Hibner U., Raibaud A., Eisen H., Baltz T., Giroud C., Baltz D. DNA rearrangements and antigenic variation in Trypanosoma equiperdum: multiple expression-linked sites in independent isolates of trypanosomes expressing the same antigen. Mol Cell Biol. 1983 Mar;3(3):399–409. doi: 10.1128/mcb.3.3.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Majiwa P. A., Young J. R., Englund P. T., Shapiro S. Z., Williams R. O. Two distinct forms of surface antigen gene rearrangement in Trypanosoma brucei. Nature. 1982 Jun 10;297(5866):514–516. doi: 10.1038/297514a0. [DOI] [PubMed] [Google Scholar]
  28. Majumder H. K., Boothroyd J. C., Weber H. Homologous 3'-terminal regions of mRNAs for surface antigens of different antigenic variants of Trypanosoma brucei. Nucleic Acids Res. 1981 Sep 25;9(18):4745–4753. doi: 10.1093/nar/9.18.4745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Matthyssens G., Michiels F., Hamers R., Pays E., Steinert M. Two variant surface glycoproteins of Trypanosoma brucei have a conserved C-terminus. Nature. 1981 Sep 17;293(5829):230–233. doi: 10.1038/293230a0. [DOI] [PubMed] [Google Scholar]
  30. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  31. Michels P. A., Liu A. Y., Bernards A., Sloof P., Van der Bijl M. M., Schinkel A. H., Menke H. H., Borst P., Veeneman G. H., Tromp M. C. Activation of the genes for variant surface glycoproteins 117 and 118 in Trypanosoma brucei. J Mol Biol. 1983 Jun 5;166(4):537–556. doi: 10.1016/s0022-2836(83)80283-6. [DOI] [PubMed] [Google Scholar]
  32. Michiels F., Matthyssens G., Kronenberger P., Pays E., Dero B., Van Assel S., Darville M., Carvador A., Steinert M., Hamers R. Gene activation and re-expression of a Trypanosoma brucei variant surface glycoprotein. EMBO J. 1983;2(7):1185–1192. doi: 10.1002/j.1460-2075.1983.tb01565.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nelson R. G., Parsons M., Barr P. J., Stuart K., Selkirk M., Agabian N. Sequences homologous to the variant antigen mRNA spliced leader are located in tandem repeats and variable orphons in trypanosoma brucei. Cell. 1983 Oct;34(3):901–909. doi: 10.1016/0092-8674(83)90547-0. [DOI] [PubMed] [Google Scholar]
  34. Nelson R. G., Parsons M., Selkirk M., Newport G., Barr P. J., Agabian N. Sequences homologous to variant antigen mRNA spliced leader in Trypanosomatidae which do not undergo antigenic variation. Nature. 1984 Apr 12;308(5960):665–667. doi: 10.1038/308665a0. [DOI] [PubMed] [Google Scholar]
  35. Overath P., Czichos J., Stock U., Nonnengaesser C. Repression of glycoprotein synthesis and release of surface coat during transformation of Trypanosoma brucei. EMBO J. 1983;2(10):1721–1728. doi: 10.1002/j.1460-2075.1983.tb01648.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Parsons M., Nelson R. G., Stuart K., Agabian N. Variant antigen genes of Trypanosoma brucei: genomic alteration of a spliced leader orphon and retention of expression-linked copies during differentiation. Proc Natl Acad Sci U S A. 1984 Feb;81(3):684–688. doi: 10.1073/pnas.81.3.684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Pays E., Delauw M. F., Laurent M., Steinert M. Possible DNA modification in GC dinucleotides of Trypanosoma brucei telomeric sequences; relationship with antigen gene transcription. Nucleic Acids Res. 1984 Jul 11;12(13):5235–5247. doi: 10.1093/nar/12.13.5235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Pays E., Delauw M. F., Van Assel S., Laurent M., Vervoort T., Van Meirvenne N., Steinert M. Modifications of a Trypanosoma b. brucei antigen gene repertoire by different DNA recombinational mechanisms. Cell. 1983 Dec;35(3 Pt 2):721–731. doi: 10.1016/0092-8674(83)90105-8. [DOI] [PubMed] [Google Scholar]
  39. Pays E., Laurent M., Delinte K., Van Meirvenne N., Steinert M. Differential size variations between transcriptionally active and inactive telomeres of Trypanosoma brucei. Nucleic Acids Res. 1983 Dec 10;11(23):8137–8147. doi: 10.1093/nar/11.23.8137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Pays E., Lheureux M., Steinert M. Structure and expression of a Trypanosoma brucei gambiense variant specific antigen gene. Nucleic Acids Res. 1982 May 25;10(10):3149–3163. doi: 10.1093/nar/10.10.3149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Pays E., Van Assel S., Laurent M., Darville M., Vervoort T., Van Meirvenne N., Steinert M. Gene conversion as a mechanism for antigenic variation in trypanosomes. Cell. 1983 Sep;34(2):371–381. doi: 10.1016/0092-8674(83)90371-9. [DOI] [PubMed] [Google Scholar]
  42. Pays E., Van Meirvenne N., Le Ray D., Steinert M. Gene duplication and transposition linked to antigenic variation in Trypanosoma brucei. Proc Natl Acad Sci U S A. 1981 May;78(5):2673–2677. doi: 10.1073/pnas.78.5.2673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Penncavage N. A., Julius M. A., Marcu K. B. An African trypanosome variant surface glycoprotein gene whose expression is not activated by duplication. Nucleic Acids Res. 1983 Dec 10;11(23):8343–8357. doi: 10.1093/nar/11.23.8343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Schibler U., Hagenbüchle O., Wellauer P. K., Pittet A. C. Two promoters of different strengths control the transcription of the mouse alpha-amylase gene Amy-1a in the parotid gland and the liver. Cell. 1983 Jun;33(2):501–508. doi: 10.1016/0092-8674(83)90431-2. [DOI] [PubMed] [Google Scholar]
  45. Shapiro S. Z., Doxsey S. J. Purification of nuclei from a flagellate protozoan, Trypanosoma brucei. Anal Biochem. 1982 Nov 15;127(1):112–115. doi: 10.1016/0003-2697(82)90152-x. [DOI] [PubMed] [Google Scholar]
  46. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Van der Ploeg L. H., Cornelissen A. W., Michels P. A., Borst P. Chromosome rearrangements in Trypanosoma brucei. Cell. 1984 Nov;39(1):213–221. doi: 10.1016/0092-8674(84)90207-1. [DOI] [PubMed] [Google Scholar]
  48. Van der Ploeg L. H., Liu A. Y., Borst P. Structure of the growing telomeres of Trypanosomes. Cell. 1984 Feb;36(2):459–468. doi: 10.1016/0092-8674(84)90239-3. [DOI] [PubMed] [Google Scholar]
  49. 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]
  50. Van der Ploeg L. H., Schwartz D. C., Cantor C. R., Borst P. Antigenic variation in Trypanosoma brucei analyzed by electrophoretic separation of chromosome-sized DNA molecules. Cell. 1984 May;37(1):77–84. doi: 10.1016/0092-8674(84)90302-7. [DOI] [PubMed] [Google Scholar]
  51. 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]
  52. Vickerman K. On the surface coat and flagellar adhesion in trypanosomes. J Cell Sci. 1969 Jul;5(1):163–193. doi: 10.1242/jcs.5.1.163. [DOI] [PubMed] [Google Scholar]
  53. Williams R. O., Young J. R., Majiwa P. A. Genomic environment of T. brucei VSG genes: presence of a minichromosome. Nature. 1982 Sep 30;299(5882):417–421. doi: 10.1038/299417a0. [DOI] [PubMed] [Google Scholar]
  54. Williams R. O., Young J. R., Majiwa P. A. Genomic rearrangements correlated with antigenic variation in Trypanosoma brucei. Nature. 1979 Dec 20;282(5741):847–849. doi: 10.1038/282847a0. [DOI] [PubMed] [Google Scholar]
  55. Wright B. M., Edwards A. J., Jones V. E. Use of a cell rupturing pump for the preparation of thymocyte subcellular fractions. J Immunol Methods. 1974 Mar;4(2):281–296. doi: 10.1016/0022-1759(74)90071-4. [DOI] [PubMed] [Google Scholar]
  56. Young J. R., Shah J. S., Matthyssens G., Williams R. O. Relationship between multiple copies of a T. brucei variable surface glycoprotein gene whose expression is not controlled by duplication. Cell. 1983 Apr;32(4):1149–1159. doi: 10.1016/0092-8674(83)90298-2. [DOI] [PubMed] [Google Scholar]

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