Participation of ezrin in bacterial uptake by trophoblast giant cells

Kenta Watanabe, Masato Tachibana, Suk Kim, Masahisa Watarai

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: Trophoblast giant (TG) cells are involved in systematic removal of bacterial pathogens from the maternal-fetal interface of the placenta. In particular, TG cells have the ability to take up extracellular antigens by active phagocytosis induced by interferon-gamma (IFN-gamma). We previously reported that heat shock cognate protein 70 (Hsc70) present on the surface of TG cells mediated the uptake of Brucella abortus. However, the mechanism of bacterial uptake by TG cells is not completely understood. Here we identified ezrin, a member of ezrin-radixin-moesin (ERM) protein family, as a molecule associated with Hsc70. Methods: Mouse TG cells were employed in all experiments, and B. abortus was used as the bacterial antigen. Confirmation of the binding capacity of ERM protein was assessed by pull-down assay and ELISA using recombinant Hsc70 and ERM proteins. Ezrin was depleted using siRNA and the depletion examined by immunoblotting or immunofluorescence staining. Results: The expression level of ezrin was higher in TG cells than in trophoblast stem (TS) cells, and ezrin knockdown TG cells showed a reduction in bacterial uptake ability. Although tyrosine phosphorylation of ezrin was not related to bacterial uptake activity, localization of Hsc70 on the membrane was affected by the depletion of ezrin in TG cells. Conclusion: Ezrin associates with Hsc70 that locates on the membrane of TG cells and participates in the bacterial uptake by TG cells.

Original languageEnglish
Article number1477
Pages (from-to)95
Number of pages1
JournalReproductive Biology and Endocrinology
Volume7
DOIs
Publication statusPublished - Sep 9 2009
Externally publishedYes

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Trophoblasts
Giant Cells
HSC70 Heat-Shock Proteins
Brucella abortus
ezrin
Bacterial Antigens
Proteins
HSP70 Heat-Shock Proteins
Membranes
Phagocytosis
Recombinant Proteins
Immunoblotting
Placenta
Small Interfering RNA
Interferon-gamma
Fluorescent Antibody Technique
Tyrosine
Stem Cells
Enzyme-Linked Immunosorbent Assay
Phosphorylation

ASJC Scopus subject areas

  • Developmental Biology
  • Endocrinology
  • Reproductive Medicine

Cite this

Participation of ezrin in bacterial uptake by trophoblast giant cells. / Watanabe, Kenta; Tachibana, Masato; Kim, Suk; Watarai, Masahisa.

In: Reproductive Biology and Endocrinology, Vol. 7, 1477, 09.09.2009, p. 95.

Research output: Contribution to journalArticle

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abstract = "Background: Trophoblast giant (TG) cells are involved in systematic removal of bacterial pathogens from the maternal-fetal interface of the placenta. In particular, TG cells have the ability to take up extracellular antigens by active phagocytosis induced by interferon-gamma (IFN-gamma). We previously reported that heat shock cognate protein 70 (Hsc70) present on the surface of TG cells mediated the uptake of Brucella abortus. However, the mechanism of bacterial uptake by TG cells is not completely understood. Here we identified ezrin, a member of ezrin-radixin-moesin (ERM) protein family, as a molecule associated with Hsc70. Methods: Mouse TG cells were employed in all experiments, and B. abortus was used as the bacterial antigen. Confirmation of the binding capacity of ERM protein was assessed by pull-down assay and ELISA using recombinant Hsc70 and ERM proteins. Ezrin was depleted using siRNA and the depletion examined by immunoblotting or immunofluorescence staining. Results: The expression level of ezrin was higher in TG cells than in trophoblast stem (TS) cells, and ezrin knockdown TG cells showed a reduction in bacterial uptake ability. Although tyrosine phosphorylation of ezrin was not related to bacterial uptake activity, localization of Hsc70 on the membrane was affected by the depletion of ezrin in TG cells. Conclusion: Ezrin associates with Hsc70 that locates on the membrane of TG cells and participates in the bacterial uptake by TG cells.",
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