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The CD44 standard/ezrin complex regulates Fas-mediated apoptosis in Jurkat cells

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Abstract

The transmembrane receptor CD44 conveys important signals from the extracellular microenvironment to the cytoplasm, a phenomena known as “outside-in” signaling. CD44 exists as several isoforms that result from alternative splicing, which differ only in the extracellular domain but yet exhibit different activities. CD44 is a binding partner for the membrane-cytoskeleton cross-linker protein ezrin. In this study, we demonstrate that only CD44 standard (CD44s) colocalizes and interacts with the actin cross-linkers ezrin and moesin using well-characterized cell lines engineered to express different CD44 isoforms. Importantly, we also show that the association CD44s-ezrin-actin is an important modulator of Fas-mediated apoptosis. The results highlight a mechanism by which signals from the extracellular milieu regulate intracellular signaling activities involved in programmed cell death.

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Acknowledgements

The authors would like to thank to Drs. Thomas Harder (Oxford, UK) for introducing the cavitation bomb method and A. Fontana (University Hospital, Zürich, Switzerland) for kindly providing us with the Neuro-2A cells expressing murine FasL. Thanks go also to Drs A. Rolink, H. Hirsch, I. Raineri for critical comments on the manuscript. We are grateful to Britt Johansson for making the transfectants as well as H. Kohler (Friedrich Miescher Institute, Basel) for sorting the transfectants. M. Wernli, V. Crotet, J. Samaridis, Dr. M. Ji and E. Kump for technical advice and assistance. Dr. P. Marbet and Dr. M. Hogan for their helpful advice concerning the confocal microscopy technique. Financial support: This work is supported by a grant from the Swiss Science Foundation (3100-067084.01) and the Swiss Cancer League (OCS 1265-08-2002).

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Author notes

  1. Ainhoa Mielgo

    Present address: Moores Cancer Center, University of California San Diego, La Jolla, CA, 92093, USA

Authors and Affiliations

  1. Institute of Medical Microbiology, Department of Clinical and Biological Sciences, University of Basel, Petersplatz 10, 4003, Basel, Switzerland

    Ainhoa Mielgo, Andrea Glaser-Ruhm, Peter Erb & Ursula Günthert

  2. Moores Cancer Center, University of California San Diego, La Jolla, CA, 92093, USA

    Dwayne Stupack

  3. Genome Center and Medical Pharmacology and Toxicology, University of California, 4513 GBSF, Davis, CA, 95616, USA

    Vincent Brondani

  4. Center of Biomedicine, Department of Clinical and Biological Sciences, University of Basel, 4003, Basel, Switzerland

    Lukas Landmann

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Correspondence to Ursula Günthert.

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ESM 5 (PDF 188 kb)

10495_2007_115_MOESM2_ESM.tif

Expression of moesin and phosphorylated Ezrin/radixin/Moesin (pERM) in Jurkat transfectants. Western blotting showing the expression of moesin and pERM in the different transfectants. (TIFF 3815 kb)

10495_2007_115_MOESM3_ESM.tif

Colocalization between CD44s and Moesin. Jurkat cells transfected with CD44s-GFP (A), CD44sΔcyt-GFP (B) and CD44v-GFP (C). Cells were stained with a mouse anti-human moesin antibody followed by a goat anti-mouse Alexa fluor 568. The merge in yellow represents colocalization between CD44s and moesin. (TIFF 31618 kb)

10495_2007_115_MOESM4_ESM.tif

Colocalization between CD44s and phosphorylated Ezrin/Moesin (pERM). Jurkat cells transfected with CD44s-GFP (A), CD44sΔcyt-GFP (B) and CD44v-GFP (C). Cells were stained with a rabbit anti-human pERM antibody followed by a goat anti-rabbit Alexa fluor 568. The merge in yellow represents colocalization between CD44s and pERM. (TIFF 30366 kb)

10495_2007_115_MOESM5_ESM.tif

Expression of CD44 isoforms and apoptosis induction on primary lymphocytes. Lymph node cells from 6 wild type and 6 CD44v6v7 KO mice were extracted and stimulated for 24, 48 and 72 hours with Phytohemagglutinine (PHA) or anti-CD3 antibody. Expression of Fas, and CD44 was measured by flow cytometry at the different time points. The apoptotic sub G1 population resulting from activation induced cell death (AICD) of lymph node cells was analyzed at 24, 48 and 72 hours by propidium iodide staining. (TIFF 14142 kb)

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Mielgo, A., Brondani, V., Landmann, L. et al. The CD44 standard/ezrin complex regulates Fas-mediated apoptosis in Jurkat cells. Apoptosis 12, 2051–2061 (2007). https://doi.org/10.1007/s10495-007-0115-3

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