Chlamydia infection of epithelial cells expressing dynamin and Eps15 mutants: Clathrin-independent entry into cells and dynamin-dependent productive growth

Haralabia Boleti*, Alexandre Benmerah, David M. Ojcius, Nadine Cerf-Bensussan, Alice Dautry-Varsat

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

79 Scopus citations

Abstract

Chlamydiae enter epithelial cells via a mechanism that still remains to be fully elucidated. In this study we investigated the pathway of entry of C. psittaci GPIC and C. trachomatis LGV/L2 into HeLa cells and demonstrated that it does not depend on clathrin coated vesicle formation. We used mutant cell lines defective in clathrin-mediated endocytosis due to overexpression of dominant negative mutants of either dynamin I or Eps15 proteins. When clathrin-dependent endocytosis was inhibited by overexpression of the dyn(K44A) mutant of dynamin I (defective in GTPase activity), Chlamydia entry was not affected. However, in these cells there was a dramatic inhibition in the proliferation of Chlamydia and the growth of the chlamydia vacuole (inclusion). When clathrin-dependent endocytosis was inhibited by overexpression of an Eps15 dominant negative mutant, the entry and growth of Chlamydia was unaltered. These results indicate that the effect on the growth of Chlamydia in the dyn(K44A) cells was not simply due to a deprivation of nutrients taken up by endocytosis. Instead, the dominant-negative mutant of dynamin most likely affects the vesicular traffic between the Chlamydia inclusion and intracellular membrane compartments. In addition, cytochalasin D inhibited Chlamydia entry by more than 90%, indicating that chlamydiae enter epithelial cells by an actin-dependent mechanism resembling phagocytosis. Finally, dynamin is apparently not involved in the formation of phagocytic vesicles containing Chlamydia.

Original languageEnglish
Pages (from-to)1487-1496
Number of pages10
JournalJournal of Cell Science
Volume112
Issue number10
StatePublished - 1999
Externally publishedYes

Keywords

  • Chlamydia
  • Dynamin
  • Endocytosis
  • Eps 15
  • Phagocytosis
  • Vesicular traffic

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