Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1990 Aug;9(8):2611–2619. doi: 10.1002/j.1460-2075.1990.tb07443.x

Architectural organization in the interphase nucleus of the protozoan Trypanosoma brucei: location of telomeres and mini-chromosomes.

H M Chung 1, C Shea 1, S Fields 1, R N Taub 1, L H Van der Ploeg 1, D B Tse 1
PMCID: PMC552294  PMID: 2369903

Abstract

We studied the spatial organization of chromatin in the interphase G1, S and G2 nucleus of the protozoan Trypanosoma brucei, applying in situ hybridization with conventional fluorescence and confocal scanning optical microscopy. The majority of the trypanosome telomere GGGTTA repeats from different chromosomes were found clustered together, either extending in a network through the nuclear interior or localized at the nuclear periphery. The population of one hundred mini-chromosomes was often asymmetrically located: either clustered in a narrow band in close association with the nuclear envelope or distributed into several clusters that segregated into roughly one half of the nucleus. The nuclear organization may undergo modifications during the cell cycle and development. We conclude that non-random spatial positioning of DNA exists in the nucleus of this protozoan. Finding a high level of structural organization in the interphase nucleus of T.brucei is an important first step towards understanding chromosome structure and functioning and its role in the control of gene expression.

Full text

PDF
2616

Images in this article

Selected References

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

  1. Agard D. A., Sedat J. W. Three-dimensional architecture of a polytene nucleus. Nature. 1983 Apr 21;302(5910):676–681. doi: 10.1038/302676a0. [DOI] [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. Berman S. A., Bursztajn S., Bowen B., Gilbert W. Localization of an acetylcholine receptor intron to the nuclear membrane. Science. 1990 Jan 12;247(4939):212–214. doi: 10.1126/science.1688472. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Borden J., Manuelidis L. Movement of the X chromosome in epilepsy. Science. 1988 Dec 23;242(4886):1687–1691. doi: 10.1126/science.3201257. [DOI] [PubMed] [Google Scholar]
  6. Borst P., van der Ploeg M., van Hoek J. F., Tas J., James J. On the DNA content and ploidy of trypanosomes. Mol Biochem Parasitol. 1982 Jul;6(1):13–23. doi: 10.1016/0166-6851(82)90049-4. [DOI] [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. Clayton C. E. The molecular biology of the Kinetoplastidae. Genet Eng. 1988;(7):1–56. [PubMed] [Google Scholar]
  9. Comings D. E. Arrangement of chromatin in the nucleus. Hum Genet. 1980 Feb;53(2):131–143. doi: 10.1007/BF00273484. [DOI] [PubMed] [Google Scholar]
  10. Cooke H. J., Smith B. A. Variability at the telomeres of the human X/Y pseudoautosomal region. Cold Spring Harb Symp Quant Biol. 1986;51(Pt 1):213–219. doi: 10.1101/sqb.1986.051.01.026. [DOI] [PubMed] [Google Scholar]
  11. Englund P. T., Hajduk S. L., Marini J. C. The molecular biology of trypanosomes. Annu Rev Biochem. 1982;51:695–726. doi: 10.1146/annurev.bi.51.070182.003403. [DOI] [PubMed] [Google Scholar]
  12. Foe V. E., Alberts B. M. Reversible chromosome condensation induced in Drosophila embryos by anoxia: visualization of interphase nuclear organization. J Cell Biol. 1985 May;100(5):1623–1636. doi: 10.1083/jcb.100.5.1623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fu X. D., Maniatis T. Factor required for mammalian spliceosome assembly is localized to discrete regions in the nucleus. Nature. 1990 Feb 1;343(6257):437–441. doi: 10.1038/343437a0. [DOI] [PubMed] [Google Scholar]
  14. Gibson W. C., Osinga K. A., Michels P. A., Borst P. Trypanosomes of subgenus Trypanozoon are diploid for housekeeping genes. Mol Biochem Parasitol. 1985 Sep;16(3):231–242. doi: 10.1016/0166-6851(85)90066-0. [DOI] [PubMed] [Google Scholar]
  15. Glass D. J., Polvere R. I., Van der Ploeg L. H. Conserved sequences and transcription of the hsp70 gene family in Trypanosoma brucei. Mol Cell Biol. 1986 Dec;6(12):4657–4666. doi: 10.1128/mcb.6.12.4657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hiraoka Y., Minden J. S., Swedlow J. R., Sedat J. W., Agard D. A. Focal points for chromosome condensation and decondensation revealed by three-dimensional in vivo time-lapse microscopy. Nature. 1989 Nov 16;342(6247):293–296. doi: 10.1038/342293a0. [DOI] [PubMed] [Google Scholar]
  17. Hochstrasser M., Sedat J. W. Three-dimensional organization of Drosophila melanogaster interphase nuclei. I. Tissue-specific aspects of polytene nuclear architecture. J Cell Biol. 1987 Jun;104(6):1455–1470. doi: 10.1083/jcb.104.6.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lawrence J. B., Singer R. H., Marselle L. M. Highly localized tracks of specific transcripts within interphase nuclei visualized by in situ hybridization. Cell. 1989 May 5;57(3):493–502. doi: 10.1016/0092-8674(89)90924-0. [DOI] [PubMed] [Google Scholar]
  19. Lawrence J. B., Villnave C. A., Singer R. H. Sensitive, high-resolution chromatin and chromosome mapping in situ: presence and orientation of two closely integrated copies of EBV in a lymphoma line. Cell. 1988 Jan 15;52(1):51–61. doi: 10.1016/0092-8674(88)90530-2. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Leon W., Frasch A. C., Hoeijmakers J. H., Fase-Fowler F., Borst P., Brunel F., Davison J. Maxi-circles and mini-circles in kinetoplast DNA from trypanosoma cruzi. Biochim Biophys Acta. 1980 Apr 30;607(2):221–231. doi: 10.1016/0005-2787(80)90075-1. [DOI] [PubMed] [Google Scholar]
  22. Mathog D., Hochstrasser M., Gruenbaum Y., Saumweber H., Sedat J. Characteristic folding pattern of polytene chromosomes in Drosophila salivary gland nuclei. 1984 Mar 29-Apr 4Nature. 308(5958):414–421. doi: 10.1038/308414a0. [DOI] [PubMed] [Google Scholar]
  23. Mathog D., Sedat J. W. The three-dimensional organization of polytene nuclei in male Drosophila melanogaster with compound XY or ring X chromosomes. Genetics. 1989 Feb;121(2):293–311. doi: 10.1093/genetics/121.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Newport J. W., Forbes D. J. The nucleus: structure, function, and dynamics. Annu Rev Biochem. 1987;56:535–565. doi: 10.1146/annurev.bi.56.070187.002535. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Sen D., Gilbert W. Formation of parallel four-stranded complexes by guanine-rich motifs in DNA and its implications for meiosis. Nature. 1988 Jul 28;334(6180):364–366. doi: 10.1038/334364a0. [DOI] [PubMed] [Google Scholar]
  27. Sloof P., Menke H. H., Caspers M. P., Borst P. Size fractionation of Trypanosoma brucei DNA: localization of the 177-bp repeat satellite DNA and a variant surface glycoprotein gene in a mini-chromosomal DNA fraction. Nucleic Acids Res. 1983 Jun 25;11(12):3889–3901. doi: 10.1093/nar/11.12.3889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Solari A. J. The 3-dimensional fine structure of the mitotic spindle in Trypanosoma cruzi. Chromosoma. 1980;78(2):239–255. doi: 10.1007/BF00328395. [DOI] [PubMed] [Google Scholar]
  29. Solari A. J. The ultrastructure of mitotic nuclei of Blastocrithidia triatomae. Z Parasitenkd. 1983;69(1):3–15. doi: 10.1007/BF00934005. [DOI] [PubMed] [Google Scholar]
  30. Sundquist W. I., Klug A. Telomeric DNA dimerizes by formation of guanine tetrads between hairpin loops. Nature. 1989 Dec 14;342(6251):825–829. doi: 10.1038/342825a0. [DOI] [PubMed] [Google Scholar]
  31. Szostak J. W., Blackburn E. H. Cloning yeast telomeres on linear plasmid vectors. Cell. 1982 May;29(1):245–255. doi: 10.1016/0092-8674(82)90109-x. [DOI] [PubMed] [Google Scholar]
  32. Tse D. B., Al-Haideri M., Pernis B., Cantor C. R., Wang C. Y. Intracellular accumulation of T-cell receptor complex molecules in a human T-cell line. Science. 1986 Nov 7;234(4777):748–751. doi: 10.1126/science.3490690. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. 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]
  35. 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]
  36. 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]
  37. Williamson J. R., Raghuraman M. K., Cech T. R. Monovalent cation-induced structure of telomeric DNA: the G-quartet model. Cell. 1989 Dec 1;59(5):871–880. doi: 10.1016/0092-8674(89)90610-7. [DOI] [PubMed] [Google Scholar]
  38. de Lange T., Shiue L., Myers R. M., Cox D. R., Naylor S. L., Killery A. M., Varmus H. E. Structure and variability of human chromosome ends. Mol Cell Biol. 1990 Feb;10(2):518–527. doi: 10.1128/mcb.10.2.518. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES