1887

Abstract

A large-scale analysis of proteins involved in host-cell signalling pathways was performed using chlamydia-infected murine cells in order to identify host proteins that are differentially activated or localized following infection. Two proteins whose distribution was altered in -infected cells relative to mock-infected cells were the actin-binding protein adducin and the regulatory kinase Raf-1. Immunoblot analysis with antibodies to both phosphorylated and non-phosphorylated forms of these proteins demonstrated that the abundance of each protein was markedly reduced in the cytosolic fraction of - and -infected cells, but the total cellular protein abundance remained unaffected by infection. Fluorescence microscopy of chlamydia-infected cells using anti--adducin antibodies demonstrated labelling at or near the chlamydial inclusion membrane. Treatment of infected cells with nocodazole or cytochalasin D did not affect -adducin that was localized to the margins of the inclusion. The demonstration of -adducin and Raf-1 redistribution within cells infected by different chlamydiae provides novel opportunities for analysis of host–pathogen interactions in this system.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.2008/020941-0
2008-12-01
2021-08-01
Loading full text...

Full text loading...

/deliver/fulltext/micro/154/12/3848.html?itemId=/content/journal/micro/10.1099/mic.0.2008/020941-0&mimeType=html&fmt=ahah

References

  1. Beatty W. L. 2006; Trafficking from CD63-positive late endocytic multivesicular bodies is essential for intracellular development of Chlamydia trachomatis . J Cell Sci 119:350–359
    [Google Scholar]
  2. Carabeo R. A., Mead D. J., Hackstadt T. 2003; Golgi-dependent transport of cholesterol to the Chlamydia trachomatis inclusion. Proc Natl Acad Sci U S A 100:6771–6776
    [Google Scholar]
  3. Geisler W. M., Suchland R. J., Rockey D. D., Stamm W. E. 2001; Epidemiology and clinical manifestations of unique Chlamydia trachomatis isolates that occupy nonfusogenic inclusions. J Infect Dis 184:879–884
    [Google Scholar]
  4. Gilligan D. M., Lozovatsky L., Gwynn B., Brugnara C., Mohandas N., Peters L. L. 1999; Targeted disruption of the beta adducin gene (Add2) causes red blood cell spherocytosis in mice. Proc Natl Acad Sci U S A 96:10717–10722
    [Google Scholar]
  5. Gilligan D. M., Sarid R., Weese J. 2002; Adducin in platelets: activation-induced phosphorylation by PKC and proteolysis by calpain. Blood 99:2418–2426
    [Google Scholar]
  6. Greene W., Zhong G. 2003; Inhibition of host cell cytokinesis by Chlamydia trachomatis infection. J Infect 47:45–51
    [Google Scholar]
  7. Grieshaber S. S., Grieshaber N. A., Hackstadt T. 2003; Chlamydia trachomatis uses host cell dynein to traffic to the microtubule-organizing centre in a p50 dynamitin-independent process. J Cell Sci 116:3793–3802
    [Google Scholar]
  8. Hackstadt T., Scidmore M. A., Rockey D. D. 1995; Lipid metabolism in Chlamydia trachomatis-infected cells: directed trafficking of Golgi-derived sphingolipids to the chlamydial inclusion. Proc Natl Acad Sci U S A 92:4877–4881
    [Google Scholar]
  9. Holleran E. A., Karki S., Holzbaur E. L. 1998; The role of the dynactin complex in intracellular motility. Int Rev Cytol 182:69–109
    [Google Scholar]
  10. Imamdi R., de Graauw M., van de Water B. 2004; Protein kinase C mediates cisplatin induced loss of adherens junctions followed by apoptosis of renal proximal tubular epithelial cells. J Pharmacol Exp Ther 311:892–903
    [Google Scholar]
  11. Jewett T. J., Fischer E. R., Mead D. J., Hackstadt T. 2006; Chlamydial TARP is a bacterial nucleator of actin. Proc Natl Acad Sci U S A 103:15599–15604
    [Google Scholar]
  12. Joshi R., Gilligan D. M., Otto E., McLaughlin T., Bennett V. 1991; Primary structure and domain organization of human alpha and beta adducin. J Cell Biol 115:665–675
    [Google Scholar]
  13. Lad S. P., Fukuda E. Y., Li J., de la Maza L. M., Li E. 2005; Up-regulation of the JAK/STAT1 signal pathway during Chlamydia trachomatis infection. J Immunol 174:7186–7193
    [Google Scholar]
  14. Larsson C. 2006; Protein kinase C and the regulation of the actin cytoskeleton. Cell Signal 18:276–284
    [Google Scholar]
  15. Matsuoka Y., Hughes C. A., Bennett V. 1996; Adducin regulation. Definition of the calmodulin-binding domain and sites of phosphorylation by protein kinases A and C. J Biol Chem 271:25157–25166
    [Google Scholar]
  16. Miyairi I., Byrne G. I. 2006; Chlamydia and programmed cell death. Curr Opin Microbiol 9:102–108
    [Google Scholar]
  17. Rockey D. D., Rosquist J. L. 1994; Protein antigens of Chlamydia psittaci present in infected cells but not detected in the infectious elementary body. Infect Immun 62:106–112
    [Google Scholar]
  18. Rockey D. D., Viratyosin W., Bannantine J. P., Suchland R. J., Stamm W. E. 2002; Diversity within inc genes of clinical Chlamydia trachomatis variant isolates that occupy non-fusogenic inclusions. Microbiology 148:2497–2505
    [Google Scholar]
  19. Rzomp K. A., Scholtes L. D., Briggs B. J., Whittaker G. R., Scidmore M. A. 2003; Rab GTPases are recruited to chlamydial inclusions in both a species-dependent and species-independent manner. Infect Immun 71:5855–5870
    [Google Scholar]
  20. Scidmore M. A., Hackstadt T. 2001; Mammalian 14-3-3 β associates with the Chlamydia trachomatis inclusion membrane via its interaction with IncG. Mol Microbiol 39:1638–1650
    [Google Scholar]
  21. Scidmore M. A., Fischer E. R., Hackstadt T. 1996a; Sphingolipids and glycoproteins are differentially trafficked to the Chlamydia trachomatis inclusion. J Cell Biol 134:363–374
    [Google Scholar]
  22. Scidmore M. A., Rockey D. D., Fischer E. R., Heinzen R. A., Hackstadt T. 1996b; Vesicular interactions of the Chlamydia trachomatis inclusion are determined by chlamydial early protein synthesis rather than route of entry. Infect Immun 64:5366–5372
    [Google Scholar]
  23. Scidmore M. A., Fischer E. R., Hackstadt T. 2003; Restricted fusion of Chlamydia trachomatis vesicles with endocytic compartments during the initial stages of infection. Infect Immun 71:973–984
    [Google Scholar]
  24. Su H., McClarty G., Dong F., Hatch G. M., Pan Z. K., Zhong G. 2004; Activation of Raf/MEK/ERK/cPLA2 signaling pathway is essential for chlamydial acquisition of host glycerophospholipids. J Biol Chem 279:9409–9416
    [Google Scholar]
  25. Suchland R. J., Rockey D. D., Bannantine J. P., Stamm W. E. 2000; Isolates of Chlamydia trachomatis that occupy nonfusogenic inclusions lack IncA, a protein localized to the inclusion membrane. Infect Immun 68:360–367
    [Google Scholar]
  26. Tamaru S., Fukuta T., Kaibuchi K., Matsuoka Y., Shiku H., Nishikawa M. 2005; Rho-kinase induces association of adducin with the cytoskeleton in platelet activation. Biochem Biophys Res Commun 332:347–351
    [Google Scholar]
  27. Valdivia R. H. 2008; Chlamydia effector proteins and new insights into chlamydial cellular microbiology. Curr Opin Microbiol 11:53–59
    [Google Scholar]
  28. Xia M., Suchland R. J., Bumgarner R. E., Peng T., Rockey D. D., Stamm W. E. 2005; Chlamydia trachomatis variant with nonfusing inclusions: growth dynamic and host-cell transcriptional response. J Infect Dis 192:1229–1236
    [Google Scholar]
  29. Yuan Y., Lyng K., Zhang Y. X., Rockey D. D., Morrison R. P. 1992; Monoclonal antibodies define genus-specific, species-specific, and cross-reactive epitopes of the chlamydial 60-kilodalton heat shock protein (hsp60): specific immunodetection and purification of chlamydial hsp60. Infect Immun 60:2288–2296
    [Google Scholar]
  30. Zhong G., Fan P., Ji H., Dong F., Huang Y. 2001; Identification of a chlamydial protease-like activity factor responsible for the degradation of host transcription factors. J Exp Med 193:935–942
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.2008/020941-0
Loading
/content/journal/micro/10.1099/mic.0.2008/020941-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error