Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1998 Jul 1;17(13):3542–3555. doi: 10.1093/emboj/17.13.3542

Elongation and clustering of glycosomes in Trypanosoma brucei overexpressing the glycosomal Pex11p.

P Lorenz 1, A G Maier 1, E Baumgart 1, R Erdmann 1, C Clayton 1
PMCID: PMC1170691  PMID: 9649425

Abstract

Kinetoplastid protozoa confine large parts of glycolysis within glycosomes, which are microbodies related to peroxisomes. We cloned the gene encoding the second most abundant integral membrane protein of Trypanosoma brucei glycosomes. The 24 kDa protein is very basic and hydrophobic, with two predicted transmembrane domains. It is targeted to peroxisomes when expressed in mammalian cells and yeast. The protein is a functional homologue of Pex11p from Saccharomyces cerevisiae: pex11Delta mutants, which are defective in peroxisome proliferation, can be complemented by the trypanosome gene. Sequence conservation is significant in the N- and C-terminal domains of all putative Pex11p homologues known, from trypanosomes, yeasts and mammals. Several lines of evidence indicate that these domains are oriented towards the cytosol. TbPex11p can form homodimers, like its yeast counterpart. The TbPEX11 gene is essential in trypanosomes. Inducible overexpression of the protein in T.brucei bloodstream forms causes growth arrest, the globular glycosomes being transformed to clusters of long tubules filling significant proportions of the cytoplasm. Reduced expression results in trypanosomes with fewer, but larger, organelles.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Aman R. A., Kenyon G. L., Wang C. C. Cross-linking of the enzymes in the glycosome of Trypanosoma brucei. J Biol Chem. 1985 Jun 10;260(11):6966–6973. [PubMed] [Google Scholar]
  3. Aman R. A., Wang C. C. Identification of two integral glycosomal membrane proteins in Trypanosoma brucei. Mol Biochem Parasitol. 1987 Aug;25(1):83–92. doi: 10.1016/0166-6851(87)90021-1. [DOI] [PubMed] [Google Scholar]
  4. Beverley S. M., Clayton C. E. Transfection of Leishmania and Trypanosoma brucei by electroporation. Methods Mol Biol. 1993;21:333–348. doi: 10.1385/0-89603-239-6:333. [DOI] [PubMed] [Google Scholar]
  5. Biebinger S., Wirtz L. E., Lorenz P., Clayton C. Vectors for inducible expression of toxic gene products in bloodstream and procyclic Trypanosoma brucei. Mol Biochem Parasitol. 1997 Mar;85(1):99–112. doi: 10.1016/s0166-6851(96)02815-0. [DOI] [PubMed] [Google Scholar]
  6. Blattner J., Dörsam H., Clatyon C. E. Function of N-terminal import signals in trypanosome microbodies. FEBS Lett. 1995 Mar 6;360(3):310–314. doi: 10.1016/0014-5793(95)00128-v. [DOI] [PubMed] [Google Scholar]
  7. Blattner J., Swinkels B., Dörsam H., Prospero T., Subramani S., Clayton C. Glycosome assembly in trypanosomes: variations in the acceptable degeneracy of a COOH-terminal microbody targeting signal. J Cell Biol. 1992 Dec;119(5):1129–1136. doi: 10.1083/jcb.119.5.1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Claros M. G., von Heijne G. TopPred II: an improved software for membrane protein structure predictions. Comput Appl Biosci. 1994 Dec;10(6):685–686. doi: 10.1093/bioinformatics/10.6.685. [DOI] [PubMed] [Google Scholar]
  9. Clayton C. E. Import of fructose bisphosphate aldolase into the glycosomes of Trypanosoma brucei. J Cell Biol. 1987 Dec;105(6 Pt 1):2649–2654. doi: 10.1083/jcb.105.6.2649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Clayton C. E., Michels P. Metabolic compartmentation in African trypanosomes. Parasitol Today. 1996 Dec;12(12):465–471. doi: 10.1016/s0169-4758(96)10073-9. [DOI] [PubMed] [Google Scholar]
  11. Dyer J. M., McNew J. A., Goodman J. M. The sorting sequence of the peroxisomal integral membrane protein PMP47 is contained within a short hydrophilic loop. J Cell Biol. 1996 Apr;133(2):269–280. doi: 10.1083/jcb.133.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Elgersma Y., Kwast L., van den Berg M., Snyder W. B., Distel B., Subramani S., Tabak H. F. Overexpression of Pex15p, a phosphorylated peroxisomal integral membrane protein required for peroxisome assembly in S.cerevisiae, causes proliferation of the endoplasmic reticulum membrane. EMBO J. 1997 Dec 15;16(24):7326–7341. doi: 10.1093/emboj/16.24.7326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Erdmann R., Blobel G. Giant peroxisomes in oleic acid-induced Saccharomyces cerevisiae lacking the peroxisomal membrane protein Pmp27p. J Cell Biol. 1995 Feb;128(4):509–523. doi: 10.1083/jcb.128.4.509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Erdmann R. The peroxisomal targeting signal of 3-oxoacyl-CoA thiolase from Saccharomyces cerevisiae. Yeast. 1994 Jul;10(7):935–944. doi: 10.1002/yea.320100708. [DOI] [PubMed] [Google Scholar]
  15. Erdmann R., Veenhuis M., Kunau W. H. Peroxisomes: Organelles at the crossroads. Trends Cell Biol. 1997 Oct;7(10):400–407. doi: 10.1016/S0962-8924(97)01126-4. [DOI] [PubMed] [Google Scholar]
  16. Evan G. I., Lewis G. K., Ramsay G., Bishop J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. doi: 10.1128/mcb.5.12.3610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Flaspohler J. A., Rickoll W. L., Beverley S. M., Parsons M. Functional identification of a Leishmania gene related to the peroxin 2 gene reveals common ancestry of glycosomes and peroxisomes. Mol Cell Biol. 1997 Mar;17(3):1093–1101. doi: 10.1128/mcb.17.3.1093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fujiki Y., Hubbard A. L., Fowler S., Lazarow P. B. Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum. J Cell Biol. 1982 Apr;93(1):97–102. doi: 10.1083/jcb.93.1.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gueiros-Filho F. J., Beverley S. M. Selection against the dihydrofolate reductase-thymidylate synthase (DHFR-TS) locus as a probe of genetic alterations in Leishmania major. Mol Cell Biol. 1996 Oct;16(10):5655–5663. doi: 10.1128/mcb.16.10.5655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Höhfeld J., Veenhuis M., Kunau W. H. PAS3, a Saccharomyces cerevisiae gene encoding a peroxisomal integral membrane protein essential for peroxisome biogenesis. J Cell Biol. 1991 Sep;114(6):1167–1178. doi: 10.1083/jcb.114.6.1167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jenö P., Mini T., Moes S., Hintermann E., Horst M. Internal sequences from proteins digested in polyacrylamide gels. Anal Biochem. 1995 Jan 1;224(1):75–82. doi: 10.1006/abio.1995.1010. [DOI] [PubMed] [Google Scholar]
  22. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  23. Marshall P. A., Dyer J. M., Quick M. E., Goodman J. M. Redox-sensitive homodimerization of Pex11p: a proposed mechanism to regulate peroxisomal division. J Cell Biol. 1996 Oct;135(1):123–137. doi: 10.1083/jcb.135.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Marshall P. A., Krimkevich Y. I., Lark R. H., Dyer J. M., Veenhuis M., Goodman J. M. Pmp27 promotes peroxisomal proliferation. J Cell Biol. 1995 Apr;129(2):345–355. doi: 10.1083/jcb.129.2.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Michels P. A., Hannaert V. The evolution of kinetoplastid glycosomes. J Bioenerg Biomembr. 1994 Apr;26(2):213–219. doi: 10.1007/BF00763070. [DOI] [PubMed] [Google Scholar]
  26. Misset O., Bos O. J., Opperdoes F. R. Glycolytic enzymes of Trypanosoma brucei. Simultaneous purification, intraglycosomal concentrations and physical properties. Eur J Biochem. 1986 Jun 2;157(2):441–453. doi: 10.1111/j.1432-1033.1986.tb09687.x. [DOI] [PubMed] [Google Scholar]
  27. Opperdoes F. R. Compartmentation of carbohydrate metabolism in trypanosomes. Annu Rev Microbiol. 1987;41:127–151. doi: 10.1146/annurev.mi.41.100187.001015. [DOI] [PubMed] [Google Scholar]
  28. Parsons M., Nielsen B. Trypanosoma brucei: two-dimensional gel analysis of the major glycosomal proteins during the life cycle. Exp Parasitol. 1990 Apr;70(3):276–285. doi: 10.1016/0014-4894(90)90109-p. [DOI] [PubMed] [Google Scholar]
  29. Passreiter M., Anton M., Lay D., Frank R., Harter C., Wieland F. T., Gorgas K., Just W. W. Peroxisome biogenesis: involvement of ARF and coatomer. J Cell Biol. 1998 Apr 20;141(2):373–383. doi: 10.1083/jcb.141.2.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rost B., Casadio R., Fariselli P., Sander C. Transmembrane helices predicted at 95% accuracy. Protein Sci. 1995 Mar;4(3):521–533. doi: 10.1002/pro.5560040318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sakai Y., Marshall P. A., Saiganji A., Takabe K., Saiki H., Kato N., Goodman J. M. The Candida boidinii peroxisomal membrane protein Pmp30 has a role in peroxisomal proliferation and is functionally homologous to Pmp27 from Saccharomyces cerevisiae. J Bacteriol. 1995 Dec;177(23):6773–6781. doi: 10.1128/jb.177.23.6773-6781.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schekman R., Orci L. Coat proteins and vesicle budding. Science. 1996 Mar 15;271(5255):1526–1533. doi: 10.1126/science.271.5255.1526. [DOI] [PubMed] [Google Scholar]
  33. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sommer J. M., Wang C. C. Targeting proteins to the glycosomes of African trypanosomes. Annu Rev Microbiol. 1994;48:105–138. doi: 10.1146/annurev.mi.48.100194.000541. [DOI] [PubMed] [Google Scholar]
  35. Subramani S., Mulligan R., Berg P. Expression of the mouse dihydrofolate reductase complementary deoxyribonucleic acid in simian virus 40 vectors. Mol Cell Biol. 1981 Sep;1(9):854–864. doi: 10.1128/mcb.1.9.854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Titorenko V. I., Ogrydziak D. M., Rachubinski R. A. Four distinct secretory pathways serve protein secretion, cell surface growth, and peroxisome biogenesis in the yeast Yarrowia lipolytica. Mol Cell Biol. 1997 Sep;17(9):5210–5226. doi: 10.1128/mcb.17.9.5210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Waterham H. R., Cregg J. M. Peroxisome biogenesis. Bioessays. 1997 Jan;19(1):57–66. doi: 10.1002/bies.950190110. [DOI] [PubMed] [Google Scholar]
  39. 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