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Molecular Architecture and Signal Transduction in Cell-to-Cell Adherens Junctions

细胞间粘附连接的分子结构和信号转导

基本信息

批准号：
03833035
负责人：
TSUKITA Sachiko
金额：
$ 1.15万
依托单位：
National Institute for Physiological Sciences
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

项目摘要

To examine the functions of ERM family members (ezrin, radixin and moesin), mouse epithelial cells (MTD-1A cells) and thymoma cells (L5178Y), which coexpress all of them, were cultured in the presence of antisense phosphorothioate oligonucleotides (PONs) complementary to ERM sequences.Immunoblotting revealed that the antisense PONs selectively suppressed the expression of each member. Immunofluorescence microscopy of these ezrin, radixin, or moesin "single-suppressed" MTD-1A cells revealed that the ERM family members are colocalized at cell-cell adhesion sites, microvilli and cleavage furrows, where actin filaments are densely associated with plasma membranes. The ezrin/radixin/moesin antisense PONs mixture induced the destruction of both cell-cell and cell-substrate adhesion and the disappearance of microvilli. Ezrin or radixin antisense PONs individually affected the initial step of the formation of both cell-cell and cell-substrate adhesion, but did not affect the microvilli structures. In sharp contrast, moesin antisense PONs did not singly affect cell-cell and cell-substrate adhesion, whereas it partly affected the microvilli structures.These data indicate that ezrin and radixin can be functionally substituted, that moesin has some synergetic functional interaction with ezrin and radixin, and that these ERM family members are involved in cell-cell and cell-substrate adhesion as well as microvilli formation.

为了研究ERM家族成员（ezrin、radixin和moesin）的功能，将它们共表达的小鼠上皮细胞（MTD-1A细胞）和胸腺瘤细胞（L5178 Y细胞）培养在与ERM序列互补的反义硫代磷酸寡核苷酸（脑桥）存在下，免疫印迹显示反义脑桥选择性抑制每个成员的表达。这些ezrin，radixin，或膜突蛋白“单抑制”MTD-1A细胞的免疫荧光显微镜显示，ERM家族成员共定位在细胞-细胞粘附位点，微绒毛和卵裂沟，其中肌动蛋白丝与质膜紧密相关。ezrin/radixin/moesin反义脑桥混合物诱导细胞-细胞和细胞-基质粘附的破坏和微绒毛的消失。Ezrin或radixin反义脑桥单独影响细胞-细胞粘附和细胞-基质粘附形成的初始步骤，但不影响微绒毛结构。与此形成鲜明对比的是，moesin反义脑桥并不单独影响细胞-细胞和细胞-基质粘附，但它部分影响微绒毛结构.这些数据表明，ezrin和radixin可以在功能上被取代，moesin与ezrin和radixin有一定的协同功能相互作用，这些ERM家族成员参与细胞-细胞和细胞-基质粘附以及微绒毛的形成。