1887

Abstract

The periodontopathic bacterium has been implicated in the pathogenesis of periodontal diseases. It has been reported previously that infection with the organism induced apoptosis in the mouse macrophage cell line J774.1. In the present study, the role of caspases during apoptosis in -infected J774.1 cells was examined. A large number of apoptotic cells was detected by flow cytometric analysis in infected J774.1 cells; however, inhibitors of caspase-9, -6 and -3/7 completely blocked the induction of apoptosis. Expression of the cleaved forms of caspase-6 and -7 was detected during apoptosis in infected J774.1 cells. Immunoblot analysis revealed that the caspase-9 inhibitor blocked expression of the cleaved forms of caspase-6 and -7, whilst the caspase-3 inhibitor blocked expression of the cleaved form of caspase-7, but not caspase-6. It is known that lamin A/C and poly(ADP-ribose) polymerase (PARP) are essential nuclear components for maintaining normal cell function and viability, and both were found to be cleaved in the infected J774.1 cells. Immunoblot analysis also revealed that the caspase-6 inhibitor blocked the cleavage of lamin A/C, whilst the caspase-3/7 inhibitor blocked the cleavage of PARP. Taken together, these results suggest that activation of caspases and the subsequent cleavage of lamin A/C and PARP are involved in the morphological changes of apoptotic macrophages infected with .

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.47193-0
2007-10-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/jmm/56/10/1399.html?itemId=/content/journal/jmm/10.1099/jmm.0.47193-0&mimeType=html&fmt=ahah

References

  1. Alnemri E. S., Livingston D. J., Nicholson D. W., Salvesen G., Thornberry N. A., Wong W. W., Yuan J. 1996; Human ICE/CED-3 protease nomenclature. Cell 87:171 [CrossRef]
    [Google Scholar]
  2. Chen Y., Smith M. R., Thirumalai K., Zychlinsky A. 1996; A bacterial invasion induces macrophage apoptosis by binding directly to ICE. EMBO J 15:3853–3860
    [Google Scholar]
  3. Christersson L. A., Wikesjö U. M. E., Aibini B., Zambon J. J., Genco R. J. 1987; Tissue localization of Actinobacillus actinomycetemcomitans in human periodontitis. II. Correlation between immunofluorescence and culture techniques. J Periodontol 58:540–545 [CrossRef]
    [Google Scholar]
  4. Cohen G. M. 1997; Caspases: the executioners of apoptosis. Biochem J 326:1–16
    [Google Scholar]
  5. Cryns V., Yuan J. 1998; Proteases to die for. Genes Dev 12:1551–1570 [CrossRef]
    [Google Scholar]
  6. Darzynkiewicz Z., Bedner E. 2000; Analysis of apoptotic cells by flow and laser scanning cytometry. Methods Enzymol 322:18–39
    [Google Scholar]
  7. Enari M., Sakahira H., Yokoyama H., Okawa K., Iwamatsu A., Nagata S. 1998; A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391:43–50 [CrossRef]
    [Google Scholar]
  8. Ghayur T., Hugunin M., Talanian R. V., Ratnofsky S., Quinlan C., Emoto Y., Pandey P., Datta R., Huang Y. other authors 1996; Proteolytic activation of protein kinase C δ by an ICE/CED 3-like protease induces characteristics of apoptosis. J Exp Med 184:2399–2404 [CrossRef]
    [Google Scholar]
  9. Goldberg M., Harel A., Gruenbaum Y. 1999; The nuclear lamina: molecular organization and interaction with chromatin. Crit Rev Eukaryot Gene Expr 9:285–293 [CrossRef]
    [Google Scholar]
  10. Gruenbaum Y., Wilson K. L., Harel A., Goldberg M., Cohen M. 2000; Review: nuclear lamins – structural proteins with fundamental functions. J Struct Biol 129:313–323 [CrossRef]
    [Google Scholar]
  11. Kaplan A. H., Weber D. J., Oddone E. Z., Perfect J. R. 1989; Infection due to Actinobacillus actinomycetemcomitans : 15 cases and review. Rev Infect Dis 11:46–63 [CrossRef]
    [Google Scholar]
  12. Kasai H., Yamamoto K., Koseki T., Yokota M., Nishihara T. 2004; Involvement of caspase activation through release of cytochrome c from mitochondria in apoptotic cell death of macrophages infected with Actinobacillus actinomycetemcomitans . FEMS Microbiol Lett 233:29–35 [CrossRef]
    [Google Scholar]
  13. Kato S., Muro M., Akifusa S., Harada N., Semba I., Fujii T., Kowashi Y., Nishihara T. 1995; Evidence for apoptosis of murine macrophages by Actinobacillus actinomycetemcomitans infection. Infect Immun 63:3914–3919
    [Google Scholar]
  14. Kaufmann S. H., Desnoyer S., Ottaviano Y., Davidson N. E., Poirier G. G. 1993; Specific proteolytic cleavage of poly(ADP-ribose) polymerase: an early marker of chemotherapy-induced apoptosis. Cancer Res 53:3976–3985
    [Google Scholar]
  15. Nicoletti I., Migliorati G., Pagliacci M. C., Grignani F., Ricardi C. A. 1991; A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139:271–279 [CrossRef]
    [Google Scholar]
  16. Nishihara T., Koseki T. 2004; Microbial etiology of periodontitis. Periodontol 2000; 36:14–26 [CrossRef]
    [Google Scholar]
  17. Nonaka K., Ishisaki A., Okahashi N., Koseki T., Kato S., Muro M., Nakashima K., Nishihara T., Kowashi Y. 2001; Involvement of caspases in apoptotic cell death of murine macrophages infected with Actinobacillus actinomycetemcomitans . J Periodontal Res 36:40–47 [CrossRef]
    [Google Scholar]
  18. Oancea M., Mazumder S., Crosby M. E., Almasan A. 2006; Apoptosis assays. Methods Mol Med 129:279–290
    [Google Scholar]
  19. Oliver F. J., de la Rubia G., Rolli V., Ruiz-Ruiz M. C., de Murcia G., Murcia J. M. 1998; Importance of poly(ADP-ribose) polymerase and its cleavage in apoptosis. Lesson from an uncleavable mutant. J Biol Chem 273:33533–33539 [CrossRef]
    [Google Scholar]
  20. Orth K., Chinnaiyan A. M., Garg M., Froelich C. J., Dixit V. M. 1996; The CED-3/ICE-like protease Mch2 is activated during apoptosis and cleaves the death substrate lamin A. J Biol Chem 271:16443–16446 [CrossRef]
    [Google Scholar]
  21. Raff M. C. 1992; Social controls on cell survival and cell death. Nature 356:397–400 [CrossRef]
    [Google Scholar]
  22. Riccardi C., Nicoletti I. 2006; Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 1:1458–1461 [CrossRef]
    [Google Scholar]
  23. Satoh M. S., Lindahl T. 1992; Role of poly(ADP-ribose) formation in DNA repair. Nature 356:356–358 [CrossRef]
    [Google Scholar]
  24. Sreenivasan P. K., Meyer D. H., Fives-Taylor P. M. 1993; Requirements for invasion of epithelial cells by Actinobacillus actinomycetemcomitans . Infect Immun 61:1239–1245
    [Google Scholar]
  25. Takahashi A., Musy P. Y., Martins L. M., Poirier G. G., Moyer R. W., Earnshaw W. C. 1996; CrmA/SPI-2 inhibition of an endogenous ICE-related protease responsible for lamin A cleavage and apoptotic nuclear fragmentation. J Biol Chem 271:32487–32490 [CrossRef]
    [Google Scholar]
  26. Thornberry N. A., Lazebnik Y. 1998; Caspases: enemies within. Science 281:1312–1316 [CrossRef]
    [Google Scholar]
  27. Yabuki M., Miyake T., Doi Y., Fujiwara T., Hamazaki K., Yoshioka T., Horton A. A., Utsumi K. 1999; Role of nuclear lamins in nuclear segmentation of human neutrophils. Physiol Chem Phys Med NMR 31:77–84
    [Google Scholar]
  28. Zhivotovsky B., Gahm A., Orrenius S. 1997; Two different proteases are involved in the proteolysis of lamin during apoptosis. Biochem Biophys Res Commun 233:96–101 [CrossRef]
    [Google Scholar]
  29. Zhuang J., Dinsdale D., Cohen G. M. 1998; Apoptosis, in human monocytic THP.1 cells, results in the release of cytochrome c from mitochondria prior to their ultracondensation, formation of outer membrane discontinuities and reduction in inner membrane potential. Cell Death Differ 5:953–962 [CrossRef]
    [Google Scholar]
  30. Zychlinsky A., Prevost M. C., Sansonetti P. J. 1992; Shigella flexneri induces apoptosis in infected macrophages. Nature 358:167–169 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.47193-0
Loading
/content/journal/jmm/10.1099/jmm.0.47193-0
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