| Project/Area Number |
03404015
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | National Institute for Physiological Sciences |
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Principal Investigator |
TSUKITA Shigenobu National Institute for Physiol. Sci. Prof., 生理学研究所, 教授 (50155347)
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| Co-Investigator(Kenkyū-buntansha) |
NAGAFUCHI Akira National Institute for Physiol. Sci. Assistant Prof., 生理学研究所, 助手 (80218023)
TSUKITA Sachiko National Institute for Physiol. Sci. Assistant Prof., 生理学研究所, 助手 (00188517)
YONEMURA Shigenobu National Institute for Physiol. Sci. Assistant Prof., 生理学研究所, 助手 (60192811)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥25,500,000 (Direct Cost: ¥25,500,000)
Fiscal Year 1993: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1992: ¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1991: ¥17,000,000 (Direct Cost: ¥17,000,000)
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| Keywords | cell adhesion / adherens junctions / radixin / undercoat / adhesion apparatus / tyrosin phosporylation / tyrosin kinase / monoclonal antibody / アドヘレンス ジャンクション / ラディキシン / 裏打ち蛋白質 / チロシリン酸化 / モノクローナル〓〓 / 細胞接着 / カドヘリン / アクチン / アンキリン / カラニン / チロシンキナ-ゼ / カテニン |
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| Research Abstract |
The ERM family members, ezrin, radixin, and moesin, localizing just beneath the plasma membranes are thought to be involved in the actin filament/plasma membrane association. To identify the intergral membrane protein directly associated with ERM family members, we performed immunoprecipitation studies using anti-moesin mAb and cultured BHK cells metabolically-labeled with [^<35>S]metionine or surface-labeled with biotin. The results indicated that moesin is directly associated with a 140kD integral membrane protein. Using BHK cells as antigens, we obtained a mAb which recognized the 140kD membrane protein. We next cloned a cDNA encoding the 140kD membrane protein and identified it as CD44, a broadly-distributed cell surface glycoprotein. Immunoprecipitation with various anti-CD44 mAbs showed that ezrin and radixin as well as moesin are associated with CD44 not only in BHK cells but also in mouse L fibroblasts. Furthermore, immunofluorescence microscopy revealed that in both BHK and L cells, the Triton X-100-insoluble CD44 is precisely colocalized with ERM family members. We concluded that ERM family members work as molecular linkers between the cytoplasmic domain of CD44 and actin-based cytoskeletons.
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