Naqvi S N, Zahn R, Mitchell D A, Stevenson B J, Munn A L
Institute of Molecular Agrobiology, National University of Singapore, Republic of Singapore.
Curr Biol. 1998 Aug 27;8(17):959-62. doi: 10.1016/s0960-9822(98)70396-3.
Several end mutations that block the internalisation step of endocytosis in Saccharomyces cerevisiae also affect the cortical actin cytoskeleton [1]. END5 encodes a proline-rich protein (End5p or verprolin) required for a polarised cortical actin cytoskeleton and endocytosis [2,3]. End5p interacts with actin [4], but its exact function is not yet known. To help elucidate End5p function, we sought other End5p-interacting proteins and identified the LAS17/BEE1 gene (encoding the yeast homologue of the human Wiskott-Aldrich Syndrome protein, WASp) as a high-copy-number suppressor of the temperature-sensitive growth and endocytic defects of end5-1 cells (carrying a frameshift mutation affecting the last 213 residues of End5p). LAS17 is unable to suppress a full deletion of END5 (end5 delta), however, suggesting that the defective End5-1p in end5-1 mutants may be stabilised by Las17p. The amino terminus of Las17p interacts with the carboxyl terminus of End5p in the yeast two-hybrid system and similar interactions have been shown between WASp and a mammalian End5p homologue, WASp-interacting protein (WIP) [5]. As las17 delta deletion mutants are blocked in endocytosis, we conclude that Las17p and End5p interact and are essential for endocytosis.
在酿酒酵母中,有几种阻断胞吞作用内化步骤的末端突变也会影响皮质肌动蛋白细胞骨架[1]。END5编码一种富含脯氨酸的蛋白质(End5p或维普洛林),它是极化皮质肌动蛋白细胞骨架和胞吞作用所必需的[2,3]。End5p与肌动蛋白相互作用[4],但其确切功能尚不清楚。为了帮助阐明End5p的功能,我们寻找了其他与End5p相互作用的蛋白质,并鉴定出LAS17/BEE1基因(编码人类威斯科特-奥尔德里奇综合征蛋白WASp的酵母同源物)是end5-1细胞(携带影响End5p最后213个残基的移码突变)温度敏感生长和胞吞缺陷的高拷贝数抑制因子。然而,LAS17无法抑制END5的完全缺失(end5 delta),这表明end5-1突变体中缺陷的End5-1p可能被Las17p稳定。在酵母双杂交系统中,Las17p的氨基末端与End5p的羧基末端相互作用,并且已显示WASp与哺乳动物End5p同源物WASp相互作用蛋白(WIP)之间存在类似的相互作用[5]。由于las17 delta缺失突变体在胞吞作用中受阻,我们得出结论,Las17p和End5p相互作用,并且对胞吞作用至关重要。