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Increase of Fas-induced apoptosis by inhibition of extracellular phosphorylation of Fas receptor in Jurkat cell line

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Abstract

Apoptosis signalling through the Fas pathway requires several steps of aggregation of the Fas receptor in the membrane, including aggregation that may occur in the absence of Fas ligand. Association of Fas domains is determinant to signal transmission following Fas ligand binding to a specific domain. The domains involved in Fas aggregation are located in its extracellular region and contain three potential protein kinase C-binding motifs. We therefore studied the possibility that phosphorylation of the extracellular region of Fas might be implicated in the regulation of Fas-mediated apoptosis. Inhibition experiments of extracellular phosphorylation were performed in human Jurkat T leukemia cells with K252b, an impermeant protein-kinase inhibitor. Extracellular phosphorylation of Fas receptor was related to ecto-kinase, as assessed by the [γ-32P] ATP labelling of Fas-116 kDa aggregates, suppressed by K252b inhibitor which significantly increased the sensitivity to Fas-mediated apoptosis. Ecto-PKC involvement was demonstrated by bisindolylmaleimide VIII, a selective inhibitor of protein kinase C which significantly increased both Fas aggregation in the membrane and Fas-mediated apoptosis and by the addition of the PKC pseudo-substrate 19–36 which inhibited the phosphorylation of 116 kDa Fas aggregates. These data support a role for Fas phosphorylation in the decreased sensitivity to apoptosis in the Jurkat T leukemia cell line.

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Authors and Affiliations

  1. Laboratory of Immunology and EA 3842, University Hospital, 87042, Limoges, France

    C. Lautrette*, E. Loum-Ribot, D. Petit & M. O. Jauberteau

  2. Diaclone Laboratories, 25020, Besançon, France

    C. Vermot-Desroches & J. Wijdenes

  3. Laboratory of Immunology, University Hospital, 2 avenue Martin Luther King, 87 042, Limoges, France

    M. O. Jauberteau

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  1. C. Lautrette*
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  2. E. Loum-Ribot
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  3. D. Petit
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  4. C. Vermot-Desroches
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  5. J. Wijdenes
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  6. M. O. Jauberteau
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Correspondence to M. O. Jauberteau.

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*There was an equal contribution from these two authors.

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Lautrette*, C., Loum-Ribot, E., Petit, D. et al. Increase of Fas-induced apoptosis by inhibition of extracellular phosphorylation of Fas receptor in Jurkat cell line. Apoptosis 11, 1195–1204 (2006). https://doi.org/10.1007/s10495-006-6795-2

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