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. 1994 Jan;62(1):126–134. doi: 10.1128/iai.62.1.126-134.1994

Selective translocation of annexins during intracellular redistribution of Chlamydia trachomatis in HeLa and McCoy cells.

M Majeed 1, J D Ernst 1, K E Magnusson 1, E Kihlström 1, O Stendahl 1
PMCID: PMC186077  PMID: 8262618

Abstract

When Chlamydia trachomatis elementary bodies enter epithelial cells, they occupy membrane-bound vesicles that aggregate with each other in a calcium-dependent manner but that do not fuse with lysosomes. As members of the annexin family of calcium- and membrane-binding proteins have been implicated in mediating calcium-regulated membrane traffic during endo- and exocytosis, we examined the intracellular localization of certain annexins following invasion of HeLa and McCoy cells by C. trachomatis serovar L2. Immunofluorescence staining with a panel of polyclonal antibodies against five human annexins revealed that annexins III, IV, and V translocate within the cytoplasm to the proximity of intracellular chlamydiae whereas the distribution of annexins I and VI was unaffected. The distinct distribution of annexins I and III was further analyzed by confocal microscopy, which revealed an intimate association between chlamydial aggregates or inclusions and annexin III. Confocal microscopy also confirmed the nonassociation of annexin I with chlamydial aggregates. Depletion of intracellular Ca2+ did not prevent association of annexin III with individual elementary body-containing endosomes but did prevent formation of chlamydial aggregates and translocation of annexin III. Furthermore, chloramphenicol-treated cells also showed association between chlamydial aggregates and annexin III, indicating that the annexins are of host cell origin. These data suggest that certain cytosolic annexins may be involved in the Ca(2+)-dependent aggregation and fusion of chlamydia-containing vesicles. The fact that these Ca(2+)-binding proteins differ in their ability to associate with chlamydia-containing vesicles and inclusions implies that the factors that regulate the interaction of annexin I and annexin III with membrane are different and suggests a selective regulatory mechanism for endosome aggregation and avoiding lysosome fusion during chlamydia infection.

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Selected References

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

  1. Blackwood R. A., Ernst J. D. Characterization of Ca2(+)-dependent phospholipid binding, vesicle aggregation and membrane fusion by annexins. Biochem J. 1990 Feb 15;266(1):195–200. doi: 10.1042/bj2660195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blyth W. A., Taverne J. Some consequences of the multiple infection of cell cultures by TRIC organisms. J Hyg (Lond) 1972 Mar;70(1):33–37. doi: 10.1017/s0022172400022063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Burgoyne R. D., Morgan A. Phospholipid-binding proteins in calcium-dependent exocytosis. Biochem Soc Trans. 1992 Nov;20(4):834–836. doi: 10.1042/bst0200834. [DOI] [PubMed] [Google Scholar]
  5. Creutz C. E., Pazoles C. J., Pollard H. B. Identification and purification of an adrenal medullary protein (synexin) that causes calcium-dependent aggregation of isolated chromaffin granules. J Biol Chem. 1978 Apr 25;253(8):2858–2866. [PubMed] [Google Scholar]
  6. Creutz C. E. The annexins and exocytosis. Science. 1992 Nov 6;258(5084):924–931. doi: 10.1126/science.1439804. [DOI] [PubMed] [Google Scholar]
  7. Crompton M. R., Moss S. E., Crumpton M. J. Diversity in the lipocortin/calpactin family. Cell. 1988 Oct 7;55(1):1–3. doi: 10.1016/0092-8674(88)90002-5. [DOI] [PubMed] [Google Scholar]
  8. Crumpton M. J., Dedman J. R. Protein terminology tangle. Nature. 1990 May 17;345(6272):212–212. doi: 10.1038/345212a0. [DOI] [PubMed] [Google Scholar]
  9. Drust D. S., Creutz C. E. Aggregation of chromaffin granules by calpactin at micromolar levels of calcium. Nature. 1988 Jan 7;331(6151):88–91. doi: 10.1038/331088a0. [DOI] [PubMed] [Google Scholar]
  10. Eissenberg L. G., Wyrick P. B., Davis C. H., Rumpp J. W. Chlamydia psittaci elementary body envelopes: ingestion and inhibition of phagolysosome fusion. Infect Immun. 1983 May;40(2):741–751. doi: 10.1128/iai.40.2.741-751.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ernst J. D. Annexin III translocates to the periphagosomal region when neutrophils ingest opsonized yeast. J Immunol. 1991 May 1;146(9):3110–3114. [PubMed] [Google Scholar]
  12. Ernst J. D., Hoye E., Blackwood R. A., Jaye D. Purification and characterization of an abundant cytosolic protein from human neutrophils that promotes Ca2(+)-dependent aggregation of isolated specific granules. J Clin Invest. 1990 Apr;85(4):1065–1071. doi: 10.1172/JCI114537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ernst J. D., Hoye E., Blackwood R. A., Mok T. L. Identification of a domain that mediates vesicle aggregation reveals functional diversity of annexin repeats. J Biol Chem. 1991 Apr 15;266(11):6670–6673. [PubMed] [Google Scholar]
  14. Fan H., Brunham R. C., McClarty G. Acquisition and synthesis of folates by obligate intracellular bacteria of the genus Chlamydia. J Clin Invest. 1992 Nov;90(5):1803–1811. doi: 10.1172/JCI116055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Friis R. R. Interaction of L cells and Chlamydia psittaci: entry of the parasite and host responses to its development. J Bacteriol. 1972 May;110(2):706–721. doi: 10.1128/jb.110.2.706-721.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fukushi H., Hirai K. Proposal of Chlamydia pecorum sp. nov. for Chlamydia strains derived from ruminants. Int J Syst Bacteriol. 1992 Apr;42(2):306–308. doi: 10.1099/00207713-42-2-306. [DOI] [PubMed] [Google Scholar]
  17. Glenney J. R., Jr Calpactins: calcium-regulated membrane-skeletal proteins. Biochem Soc Trans. 1987 Oct;15(5):798–800. doi: 10.1042/bst0150798. [DOI] [PubMed] [Google Scholar]
  18. Grayston J. T., Campbell L. A., Kuo C. C., Mordhorst C. H., Saikku P., Thom D. H., Wang S. P. A new respiratory tract pathogen: Chlamydia pneumoniae strain TWAR. J Infect Dis. 1990 Apr;161(4):618–625. doi: 10.1093/infdis/161.4.618. [DOI] [PubMed] [Google Scholar]
  19. Hatch T. P., Vance D. W., Jr, Al-Hossainy E. Attachment of Chlamydia psittaci to formaldehyde-fixed and unfixed L cells. J Gen Microbiol. 1981 Aug;125(2):273–283. doi: 10.1099/00221287-125-2-273. [DOI] [PubMed] [Google Scholar]
  20. Hodinka R. L., Davis C. H., Choong J., Wyrick P. B. Ultrastructural study of endocytosis of Chlamydia trachomatis by McCoy cells. Infect Immun. 1988 Jun;56(6):1456–1463. doi: 10.1128/iai.56.6.1456-1463.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jaconi M. E., Lew D. P., Carpentier J. L., Magnusson K. E., Sjögren M., Stendahl O. Cytosolic free calcium elevation mediates the phagosome-lysosome fusion during phagocytosis in human neutrophils. J Cell Biol. 1990 May;110(5):1555–1564. doi: 10.1083/jcb.110.5.1555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Lew D. P. Receptor signalling and intracellular calcium in neutrophil activation. Eur J Clin Invest. 1989 Aug;19(4):338–346. doi: 10.1111/j.1365-2362.1989.tb00240.x. [DOI] [PubMed] [Google Scholar]
  24. Lew P. D., Wollheim C. B., Waldvogel F. A., Pozzan T. Modulation of cytosolic-free calcium transients by changes in intracellular calcium-buffering capacity: correlation with exocytosis and O2-production in human neutrophils. J Cell Biol. 1984 Oct;99(4 Pt 1):1212–1220. doi: 10.1083/jcb.99.4.1212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Majeed M., Gustafsson M., Kihlström E., Stendahl O. Roles of Ca2+ and F-actin in intracellular aggregation of Chlamydia trachomatis in eucaryotic cells. Infect Immun. 1993 Apr;61(4):1406–1414. doi: 10.1128/iai.61.4.1406-1414.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Majeed M., Kihlström E. Mobilization of F-actin and clathrin during redistribution of Chlamydia trachomatis to an intracellular site in eucaryotic cells. Infect Immun. 1991 Dec;59(12):4465–4472. doi: 10.1128/iai.59.12.4465-4472.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moulder J. W. Comparative biology of intracellular parasitism. Microbiol Rev. 1985 Sep;49(3):298–337. doi: 10.1128/mr.49.3.298-337.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moulder J. W. Interaction of chlamydiae and host cells in vitro. Microbiol Rev. 1991 Mar;55(1):143–190. doi: 10.1128/mr.55.1.143-190.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ridderhof J. C., Barnes R. C. Fusion of inclusions following superinfection of HeLa cells by two serovars of Chlamydia trachomatis. Infect Immun. 1989 Oct;57(10):3189–3193. doi: 10.1128/iai.57.10.3189-3193.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Saikku P., Wang S. P., Kleemola M., Brander E., Rusanen E., Grayston J. T. An epidemic of mild pneumonia due to an unusual strain of Chlamydia psittaci. J Infect Dis. 1985 May;151(5):832–839. doi: 10.1093/infdis/151.5.832. [DOI] [PubMed] [Google Scholar]
  31. Sneddon J. M., Wenman W. M. The effect of ions on the adhesion and internalization of Chlamydia trachomatis by HeLa cells. Can J Microbiol. 1985 Apr;31(4):371–374. doi: 10.1139/m85-071. [DOI] [PubMed] [Google Scholar]
  32. Stephens R. S. Challenge of Chlamydia research. Infect Agents Dis. 1992 Dec;1(6):279–293. [PubMed] [Google Scholar]
  33. Söderlund G., Kihlström E. Physicochemical surface properties of elementary bodies from different serotypes of chlamydia trachomatis and their interaction with mouse fibroblasts. Infect Immun. 1982 Jun;36(3):893–899. doi: 10.1128/iai.36.3.893-899.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tauber A. I., Pavlotsky N., Lin J. S., Rice P. A. Inhibition of human neutrophil NADPH oxidase by Chlamydia serovars E, K, and L2. Infect Immun. 1989 Apr;57(4):1108–1112. doi: 10.1128/iai.57.4.1108-1112.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ward M. E., Murray A. Control mechanisms governing the infectivity of Chlamydia trachomatis for HeLa cells: mechanisms of endocytosis. J Gen Microbiol. 1984 Jul;130(7):1765–1780. doi: 10.1099/00221287-130-7-1765. [DOI] [PubMed] [Google Scholar]
  36. Wyrick P. B., Brownridge E. A. Growth of Chlamydia psittaci in macrophages. Infect Immun. 1978 Mar;19(3):1054–1060. doi: 10.1128/iai.19.3.1054-1060.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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