Maldonado-Báez Lymarie, Dores Michael R, Perkins Edward M, Drivas Theodore G, Hicke Linda, Wendland Beverly
Department of Biology and Integrated Imaging Center, The Johns Hopkins University, Baltimore, MD 21218, USA.
Mol Biol Cell. 2008 Jul;19(7):2936-48. doi: 10.1091/mbc.e07-10-1019. Epub 2008 Apr 30.
The spatial and temporal regulation of the interactions among the approximately 60 proteins required for endocytosis is under active investigation in many laboratories. We have identified the interaction between monomeric clathrin adaptors and endocytic scaffold proteins as a critical prerequisite for the recruitment and/or spatiotemporal dynamics of endocytic proteins at early and late stages of internalization. Quadruple deletion yeast cells (DeltaDeltaDeltaDelta) lacking four putative adaptors, Ent1/2 and Yap1801/2 (homologues of epsin and AP180/CALM proteins), with a plasmid encoding Ent1 or Yap1802 mutants, have defects in endocytosis and growth at 37 degrees C. Live-cell imaging revealed that the dynamics of the early- and late-acting scaffold proteins Ede1 and Pan1, respectively, depend upon adaptor interactions mediated by adaptor asparagine-proline-phenylalanine motifs binding to scaffold Eps15 homology domains. These results suggest that adaptor/scaffold interactions regulate transitions from early to late events and that clathrin adaptor/scaffold protein interaction is essential for clathrin-mediated endocytosis.
内吞作用所需的约60种蛋白质之间相互作用的时空调节正在许多实验室中积极研究。我们已经确定单体网格蛋白衔接蛋白与内吞支架蛋白之间的相互作用是内化早期和晚期内吞蛋白募集和/或时空动态的关键先决条件。缺乏四种假定衔接蛋白Ent1/2和Yap1801/2（epsin和AP180/CALM蛋白的同源物）的四重缺失酵母细胞（DeltaDeltaDeltaDelta），携带编码Ent1或Yap1802突变体的质粒，在37℃下内吞作用和生长存在缺陷。活细胞成像显示，早期和晚期作用的支架蛋白Ede1和Pan1的动态分别取决于由衔接蛋白天冬酰胺-脯氨酸-苯丙氨酸基序与支架Eps15同源结构域结合介导的衔接蛋白相互作用。这些结果表明，衔接蛋白/支架相互作用调节从早期到晚期事件的转变，并且网格蛋白衔接蛋白/支架蛋白相互作用对于网格蛋白介导的内吞作用至关重要。
