Pai L M, Kirkpatrick C, Blanton J, Oda H, Takeichi M, Peifer M
Department of Biology, University of North Carolina, Chapel Hill, North Carolina 27599-3280, USA.
J Biol Chem. 1996 Dec 13;271(50):32411-20. doi: 10.1074/jbc.271.50.32411.
Adherens junctions are multiprotein complexes mediating cell-cell adhesion and communication. They are organized around a transmembrane cadherin, which binds a set of cytoplasmic proteins required for adhesion and to link the complex to the actin cytoskeleton. Three components of Drosophila adherens junctions, analogous to those in vertebrates, have been identified: Armadillo (homolog of beta-catenin), Drosophila E-cadherin (DE-cadherin), and alpha-catenin. We carried out the first analysis of the interactions between these proteins using in vitro binding assays, the yeast two-hybrid system, and in vivo assays. We identified a 76-amino acid region of Armadillo that is necessary and sufficient for binding alpha-catenin and found that the N-terminal 258 amino acids of alpha-catenin interact with Armadillo. A large region of Armadillo, spanning six central Armadillo repeats, is required for DE-cadherin binding, whereas only 41 amino acids of the DE-cadherin cytoplasmic tail are sufficient for Armadillo binding. Our data complement and extend results obtained in studies of vertebrate adherens junctions, providing a foundation for understanding how junctional proteins assemble and a basis for interpreting existing mutations and creating new ones.
黏着连接是介导细胞间黏附和通讯的多蛋白复合体。它们围绕跨膜钙黏蛋白组织形成,该钙黏蛋白结合一组黏附所需的胞质蛋白,并将复合体与肌动蛋白细胞骨架相连。已鉴定出果蝇黏着连接的三个与脊椎动物中类似的组分:犰狳蛋白(β-连环蛋白的同源物)、果蝇E-钙黏蛋白(DE-钙黏蛋白)和α-连环蛋白。我们使用体外结合试验、酵母双杂交系统和体内试验对这些蛋白之间的相互作用进行了首次分析。我们鉴定出犰狳蛋白的一个76个氨基酸的区域,该区域对于结合α-连环蛋白是必需且足够的,并发现α-连环蛋白的N端258个氨基酸与犰狳蛋白相互作用。犰狳蛋白的一个跨越六个中央犰狳重复序列的大区域是DE-钙黏蛋白结合所必需的,而DE-钙黏蛋白胞质尾仅41个氨基酸就足以与犰狳蛋白结合。我们的数据补充并扩展了在脊椎动物黏着连接研究中获得的结果,为理解连接蛋白如何组装提供了基础,并为解释现有突变和创造新突变提供了依据。