Engagement of the Fas receptor (CD95) initiates multiple signaling pathways that lead to apoptosis, such as the formation of death-inducing signaling complex (DISC), activation of caspase cascades, and the generation of the lipid messenger, ceramide. Sphingomyelin (SM) is a major component of lipid rafts, which are specialized structures that enhance the efficiency of membrane receptor signaling and are a main source of ceramide. However, the functions of SM in Fas-mediated apoptosis have yet to be clearly defined, as the responsible genes have not been identified. After cloning a gene responsible for SM synthesis, SMS1, we established SM synthase–defective WR19L cells transfected with the human Fas gene (WR/Fas-SM(−)), and cells that have been functionally restored by transfection with SMS1 (WR/Fas-SMS1). We show that expression of membrane SM enhances Fas-mediated apoptosis through increasing DISC formation, activation of caspases, efficient translocation of Fas into lipid rafts, and subsequent Fas clustering. Furthermore, WR/Fas-SMS1 cells, but not WR/Fas-SM(−) cells, showed a considerable increase in ceramide generation within lipid rafts upon Fas stimulation. These data suggest that a membrane SM is important for Fas clustering through aggregation of lipid rafts, leading to Fas-mediated apoptosis.
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18 July 2005
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July 11 2005
Role of membrane sphingomyelin and ceramide in platform formation for Fas-mediated apoptosis
Michihiko Miyaji,
Michihiko Miyaji
1Department of Rheumatology and Clinical Immunology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
2First Department of Internal Medicine, Kansai Medical University, Moriguchi, Osaka 570-8506, Japan
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Zhe-Xiong Jin,
Zhe-Xiong Jin
1Department of Rheumatology and Clinical Immunology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
3Division of Hematology and Immunology, Department of Internal Medicine, Kanazawa Medical University, Ishikawa 920-0293, Japan
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Shohei Yamaoka,
Shohei Yamaoka
4Department of Clinical Laboratory Medicine/Hematology, Faculty of Medicine, Tottori University, Yonago, Tottori 683-8504, Japan
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Ryuichi Amakawa,
Ryuichi Amakawa
2First Department of Internal Medicine, Kansai Medical University, Moriguchi, Osaka 570-8506, Japan
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Shirou Fukuhara,
Shirou Fukuhara
2First Department of Internal Medicine, Kansai Medical University, Moriguchi, Osaka 570-8506, Japan
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Satoshi B. Sato,
Satoshi B. Sato
5RIKEN, Wako, Saitama 351-0198, Japan
6Department of Biophysics, Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan
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Toshihide Kobayashi,
Toshihide Kobayashi
5RIKEN, Wako, Saitama 351-0198, Japan
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Naochika Domae,
Naochika Domae
7Department of Medicine, Osaka Dental University, Hirakata, Osaka 573-1121, Japan
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Tsuneyo Mimori,
Tsuneyo Mimori
1Department of Rheumatology and Clinical Immunology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
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Eda T. Bloom,
Eda T. Bloom
8Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892
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Toshiro Okazaki,
Toshiro Okazaki
4Department of Clinical Laboratory Medicine/Hematology, Faculty of Medicine, Tottori University, Yonago, Tottori 683-8504, Japan
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Hisanori Umehara
Hisanori Umehara
1Department of Rheumatology and Clinical Immunology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
3Division of Hematology and Immunology, Department of Internal Medicine, Kanazawa Medical University, Ishikawa 920-0293, Japan
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Michihiko Miyaji
1Department of Rheumatology and Clinical Immunology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
2First Department of Internal Medicine, Kansai Medical University, Moriguchi, Osaka 570-8506, Japan
Zhe-Xiong Jin
1Department of Rheumatology and Clinical Immunology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
3Division of Hematology and Immunology, Department of Internal Medicine, Kanazawa Medical University, Ishikawa 920-0293, Japan
Shohei Yamaoka
4Department of Clinical Laboratory Medicine/Hematology, Faculty of Medicine, Tottori University, Yonago, Tottori 683-8504, Japan
Ryuichi Amakawa
2First Department of Internal Medicine, Kansai Medical University, Moriguchi, Osaka 570-8506, Japan
Shirou Fukuhara
2First Department of Internal Medicine, Kansai Medical University, Moriguchi, Osaka 570-8506, Japan
Satoshi B. Sato
5RIKEN, Wako, Saitama 351-0198, Japan
6Department of Biophysics, Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan
Toshihide Kobayashi
5RIKEN, Wako, Saitama 351-0198, Japan
Naochika Domae
7Department of Medicine, Osaka Dental University, Hirakata, Osaka 573-1121, Japan
Tsuneyo Mimori
1Department of Rheumatology and Clinical Immunology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
Eda T. Bloom
8Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892
Toshiro Okazaki
4Department of Clinical Laboratory Medicine/Hematology, Faculty of Medicine, Tottori University, Yonago, Tottori 683-8504, Japan
Hisanori Umehara
1Department of Rheumatology and Clinical Immunology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
3Division of Hematology and Immunology, Department of Internal Medicine, Kanazawa Medical University, Ishikawa 920-0293, Japan
CORRESPONDENCE Hisanori Umehara: [email protected]
Abbreviations used: aSMase; acid sphingomyelinase, CTx; choleratoxin B; ΔΨm, mitochondrial membrane potential; DISC, death-inducing signaling complex; FADD, Fas-associated death domain; FasL, Fas ligand; MBP, maltose-binding protein; PI, propidium iodide; SM; sphingomyelin.
Received:
August 19 2004
Accepted:
May 02 2005
Online ISSN: 1540-9538
Print ISSN: 0022-1007
The Rockefeller University Press
2005
J Exp Med (2005) 202 (2): 249–259.
Article history
Received:
August 19 2004
Accepted:
May 02 2005
Citation
Michihiko Miyaji, Zhe-Xiong Jin, Shohei Yamaoka, Ryuichi Amakawa, Shirou Fukuhara, Satoshi B. Sato, Toshihide Kobayashi, Naochika Domae, Tsuneyo Mimori, Eda T. Bloom, Toshiro Okazaki, Hisanori Umehara; Role of membrane sphingomyelin and ceramide in platform formation for Fas-mediated apoptosis . J Exp Med 18 July 2005; 202 (2): 249–259. doi: https://doi.org/10.1084/jem.20041685
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