Konzok A, Weber I, Simmeth E, Hacker U, Maniak M, Müller-Taubenberger A
Max-Planck-Institut für Biochemie, D-82152 Martinsried, Germany.
J Cell Biol. 1999 Jul 26;146(2):453-64. doi: 10.1083/jcb.146.2.453.
The 64-kD protein DAip1 from Dictyostelium contains nine WD40-repeats and is homologous to the actin-interacting protein 1, Aip1p, from Saccharomyces cerevisiae, and to related proteins from Caenorhabditis, Physarum, and higher eukaryotes. We show that DAip1 is localized to dynamic regions of the cell cortex that are enriched in filamentous actin: phagocytic cups, macropinosomes, lamellipodia, and other pseudopodia. In cells expressing green fluorescent protein (GFP)-tagged DAip1, the protein rapidly redistributes into newly formed cortical protrusions. Functions of DAip1 in vivo were assessed using null mutants generated by gene replacement, and by overexpressing DAip1. DAip1-null cells are impaired in growth and their rates of fluid-phase uptake, phagocytosis, and movement are reduced in comparison to wild-type rates. Cytokinesis is prolonged in DAip1-null cells and they tend to become multinucleate. On the basis of similar results obtained by DAip1 overexpression and effects of latrunculin-A treatment, we propose a function for DAip1 in the control of actin depolymerization in vivo, probably through interaction with cofilin. Our data suggest that DAip1 plays an important regulatory role in the rapid remodeling of the cortical actin meshwork.
盘基网柄菌的64-kD蛋白DAip1含有9个WD40重复序列,与酿酒酵母的肌动蛋白相互作用蛋白1(Aip1p)以及秀丽隐杆线虫、绒泡菌和高等真核生物的相关蛋白同源。我们发现DAip1定位于富含丝状肌动蛋白的细胞皮层动态区域:吞噬杯、大胞饮体、片状伪足和其他伪足。在表达绿色荧光蛋白(GFP)标记的DAip1的细胞中,该蛋白迅速重新分布到新形成的皮层突起中。使用基因替换产生的缺失突变体并通过过表达DAip1来评估DAip1在体内的功能。与野生型相比,DAip1缺失的细胞生长受损,其液相摄取、吞噬作用和运动速率降低。DAip1缺失的细胞中胞质分裂延长,并且它们倾向于变成多核。基于DAip1过表达获得的类似结果以及拉春库林-A处理的影响,我们提出DAip1在体内控制肌动蛋白解聚中起作用,可能是通过与丝切蛋白相互作用。我们的数据表明DAip1在皮层肌动蛋白网络的快速重塑中起重要的调节作用。