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. 1976 Mar;73(3):852–856. doi: 10.1073/pnas.73.3.852

Surface membrane carbohydrate alterations of a flagellated protozoan mediated by bacterial endosymbiotes.

D M Dwyer, K P Chang
PMCID: PMC336017  PMID: 1062797

Abstract

Crithidia oncopelti, a parasitic trypanosomatid protozoan of insects, normally contains intracellular symbiotic bacteria. As shown earlier, the protozoa can be rid of their endosymbiotes by chloramphenicol, producing a symbiote-free cell line. Here surface-membrane carbohydrate ligands of the symbiote-containing and symbiote-free strains were compared by lectin-mediated agglutination, lectin-ultrastructure localization. [3H] lectin-binding, and fluorescent lectin staining. Symbiote-free organisms consistently had 3-fold higher agglutination titers than symbiote-containing cells with concanavalin A. Conversely, symbiote-containing flagellates had 2- to 3-fold greater agglutination titers with a fucose-binding lectin than symbiote-free organisms. Ultrastructure results showed that more of concanavalin A-horseradish peroxidase-diaminobenzidine reaction product was present at the surface of symbiote-free than on symbiote-containing cells. Treatment with [3H]concanavalin A revealed that surface membrane sites available per cell for [3H]lectin-binding ranged from 6.2 to 7.4 x 10(4) and from 24 to 27 x 10(4) for symbiote-containing and symbiote-free organisms, respectively, i.e., the mean binding level of the latter for the lectin was 3.5 times greater than that of the former. Moreover, symbiote-free cells fluoresced more than symbiote-containing organisms after staining with fluorescein-labeled concanavalin A. Apparently, the prokaryotic endosymbiotes somehow alter the quantity of saccharide ligands in the C. oncopelti surface membrane.

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Selected References

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  1. Alves M. J., Colli W. Agglutination of Trypanosoma cruzi by concanavalin A. J Protozool. 1974 Oct;21(4):575–578. doi: 10.1111/j.1550-7408.1974.tb03704.x. [DOI] [PubMed] [Google Scholar]
  2. Bernhard W., Avrameas S. Ultrastructural visualization of cellular carbohydrate components by means of concanavalin A. Exp Cell Res. 1971 Jan;64(1):232–236. doi: 10.1016/0014-4827(71)90217-5. [DOI] [PubMed] [Google Scholar]
  3. Burger M. M. A difference in the architecture of the surface membrane of normal and virally transformed cells. Proc Natl Acad Sci U S A. 1969 Mar;62(3):994–1001. doi: 10.1073/pnas.62.3.994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chang K. P., Chang C. S., Sassa S. Heme biosynthesis in bacterium-protozoon symbioses: enzymic defects in host hemoflagellates and complemental role of their intracellular symbiotes. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2979–2983. doi: 10.1073/pnas.72.8.2979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang K. P. Haematophagous insect and haemoflagellate as hosts for prokaryotic endosymbionts. Symp Soc Exp Biol. 1975;(29):407–428. [PubMed] [Google Scholar]
  6. Chang K. P. Reduced growth of Blastocrithidia culicis and Crithidia oncopelti freed of intracellular symbiotes by chloramphenicol. J Protozool. 1975 May;22(2):271–276. doi: 10.1111/j.1550-7408.1975.tb05866.x. [DOI] [PubMed] [Google Scholar]
  7. Chang K. P., Trager W. Nutritional significance of symbiotic bacteria in two species of hemoflagellates. Science. 1974 Feb 8;183(4124):531–532. doi: 10.1126/science.183.4124.531. [DOI] [PubMed] [Google Scholar]
  8. Chang K. P. Ultrastructure of symbiotic bacteria in normal and antibiotic-treated Blastocrithidia culicis and Crithidia oncopelti. J Protozool. 1974 Nov;21(5):699–707. doi: 10.1111/j.1550-7408.1974.tb03733.x. [DOI] [PubMed] [Google Scholar]
  9. Collard J. G., Temmink J. H. Differences in density of Concanavalin A-binding sites due to differences in surface morphology of suspended normal and transformed 3T3 fibroblasts. J Cell Sci. 1975 Oct;19(1):21–32. doi: 10.1242/jcs.19.1.21. [DOI] [PubMed] [Google Scholar]
  10. Drlica K. A., Kado C. I. Crown gall tumors: are bacterial nucleic acids involved? Bacteriol Rev. 1975 Sep;39(3):186–196. doi: 10.1128/br.39.3.186-196.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dwyer D. M. Lectin binding saccharides on a parasitic protozoan. Science. 1974 Apr 26;184(4135):471–473. doi: 10.1126/science.184.4135.471. [DOI] [PubMed] [Google Scholar]
  12. Gibson D. A., Marquardt M. D., Gordon J. A. Cell rigidity: Effect on concanavalin A-mediated agglutinability of fibroblasts after fixation. Science. 1975 Jul 4;189(4196):45–46. doi: 10.1126/science.166434. [DOI] [PubMed] [Google Scholar]
  13. Kaneko I., Hayatsu H., Ukita T. A quantitative assay for concanavalin A- and Ricinus communis agglutinin-mediated agglutinations of rat ascites hepatoma cells. Relationship between concanavalin A binding and cell agglutination. Biochim Biophys Acta. 1975 May 5;392(1):131–140. doi: 10.1016/0304-4165(75)90173-7. [DOI] [PubMed] [Google Scholar]
  14. Knopf U. C. On a eukaryotic cell-transformation by a bacterium. Subcell Biochem. 1974 Mar;3(1):39–48. [PubMed] [Google Scholar]
  15. MORGAN J. F., MORTON H. J., PARKER R. C. Nutrition of animal cells in tissue culture; initial studies on a synthetic medium. Proc Soc Exp Biol Med. 1950 Jan;73(1):1–8. doi: 10.3181/00379727-73-17557. [DOI] [PubMed] [Google Scholar]
  16. Marquardt M. D., Gordon J. A. Glutaraldehyde fixation and the mechanism of erythroycte agglutination by concanavalin A and soybean agglutinin. Exp Cell Res. 1975 Mar 15;91(2):310–316. doi: 10.1016/0014-4827(75)90109-3. [DOI] [PubMed] [Google Scholar]
  17. Martin B. J., Spicer S. S. Letter: Concanavalin A-iron dextran technique for staining cell surface mucosubstances. J Histochem Cytochem. 1974 Mar;22(3):206–207. doi: 10.1177/22.3.206. [DOI] [PubMed] [Google Scholar]
  18. McCracken R. O., Lumsden R. D. Structure and function of parasite surface membranes. II. Concanavalin A adsorption by the cestode Hymenolepis diminuta and its effect on transport. Comp Biochem Physiol B. 1975 Oct 15;52(2):331–337. doi: 10.1016/0305-0491(75)90074-7. [DOI] [PubMed] [Google Scholar]
  19. Mundim M. H., Roitman I., Hermans M. A., Kitajima E. W. Simple nutrition of Crithidia deanei, a reduviid trypanosomatid with an endosymbiont. J Protozool. 1974 Oct;21(4):518–521. doi: 10.1111/j.1550-7408.1974.tb03691.x. [DOI] [PubMed] [Google Scholar]
  20. NEWTON B. A., HORNE R. W. Intracellular structures in Strigomonas oncopelti. I. Cytoplasmic structures containing ribonucleoprotein. Exp Cell Res. 1957 Dec;13(3):563–574. doi: 10.1016/0014-4827(57)90086-1. [DOI] [PubMed] [Google Scholar]
  21. Nicolson G. L., Lacorbiere M. Cell contact-dependent increase in membrane D-galactopyranosyl-like residues on normal, but not virus- or spontaneously-transformed, murine fibroblasts. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1672–1676. doi: 10.1073/pnas.70.6.1672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nicolson G. L., Lacorbiere M., Eckhart W. Qualitative and quantitative interactions of lectins with untreated and neuraminidase-treated normal, wild-type, and temperature-sensitive polyoma-transformed fibroblasts. Biochemistry. 1975 Jan 14;14(1):172–179. doi: 10.1021/bi00672a029. [DOI] [PubMed] [Google Scholar]
  23. Nicolson G. L. Neuraminidase "unmasking" and failure of trypsin to "unmask" -D-galactose-like sites on erythrocyte, lymphoma, and normal and virus-transformed fibroblast cell membranes. J Natl Cancer Inst. 1973 Jun;50(6):1443–1451. doi: 10.1093/jnci/50.6.1443. [DOI] [PubMed] [Google Scholar]
  24. Nicolson G. L. The interactions of lectins with animal cell surfaces. Int Rev Cytol. 1974;39:89–190. doi: 10.1016/s0074-7696(08)60939-0. [DOI] [PubMed] [Google Scholar]
  25. Preer J. R., Jr, Preer L. B., Jurand A. Kappa and other endosymbionts in Paramecium aurelia. Bacteriol Rev. 1974 Jun;38(2):113–163. doi: 10.1128/br.38.2.113-163.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rapin A. M., Burger M. M. Tumor cell surfaces: general alterations detected by agglutinins. Adv Cancer Res. 1974;20:1–91. doi: 10.1016/s0065-230x(08)60108-6. [DOI] [PubMed] [Google Scholar]
  27. Sachs L. Regulation of membrane changes, differentiation, and malignancy in carcinogenesis. Harvey Lect. 1974;68:1–35. [PubMed] [Google Scholar]
  28. Spencer R., Cross G. A. Purification and properties of nucleic acids from an unusual cytoplasmic organelle in the flagellate protozoan Crithidia oncopelti. Biochim Biophys Acta. 1975 May 1;390(2):141–154. doi: 10.1016/0005-2787(75)90337-8. [DOI] [PubMed] [Google Scholar]
  29. Tuan R. S., Chang K. P. Isolation of intracellular symbiotes by immune lysis of flagellate protozoa and characterization of their DNA. J Cell Biol. 1975 May;65(2):309–323. doi: 10.1083/jcb.65.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weiss E. Growth and physiology of rickettsiae. Bacteriol Rev. 1973 Sep;37(3):259–283. doi: 10.1128/br.37.3.259-283.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. van den Berg K. J., Betel I. Increased transport of 2-aminoisobutyric acid in rat lymphocytes stimulated with concanavalin A. Exp Cell Res. 1973 Jan;76(1):63–72. doi: 10.1016/0014-4827(73)90419-9. [DOI] [PubMed] [Google Scholar]

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