Sehring Ivonne M, Reiner Christoph, Mansfeld Jörg, Plattner Helmut, Kissmehl Roland
Department of Biology, University of Konstanz, P.O. Box 5560, 78457 Konstanz, Germany.
J Cell Sci. 2007 Jan 1;120(Pt 1):177-90. doi: 10.1242/jcs.03313. Epub 2006 Dec 12.
To localize the different actin paralogs found in Paramecium and to disclose functional implications, we used overexpression of GFP-fusion proteins and antibody labeling, as well as gene silencing. Several isoforms are associated with food vacuoles of different stages. GFP-actin either forms a tail at the lee side of the organelle, or it is vesicle bound in a homogenous or in a speckled arrangement, thus reflecting an actin-based mosaic of the phagosome surface appropriate for association and/or dissociation of other vesicles upon travel through the cell. Several paralogs occur in cilia. A set of actins is found in the cell cortex where actin outlines the regular surface pattern. Labeling of defined structures of the oral cavity is due to other types of actin, whereas yet more types are distributed in a pattern suggesting association with the numerous Golgi fields. A substantial fraction of actins is associated with cytoskeletal elements that are known to be composed of other proteins. Silencing of the respective actin genes or gene subfamilies entails inhibitory effects on organelles compatible with localization studies. Knock down of the actin found in the cleavage furrow abolishes cell division, whereas silencing of other actin genes alters vitality, cell shape and swimming behavior.
为了定位草履虫中发现的不同肌动蛋白旁系同源物并揭示其功能意义,我们使用了绿色荧光蛋白(GFP)融合蛋白的过表达、抗体标记以及基因沉默技术。几种亚型与不同阶段的食物泡相关。GFP-肌动蛋白要么在细胞器的背侧形成一条尾巴,要么以均匀或斑点状排列与囊泡结合,从而反映出吞噬体表面基于肌动蛋白的镶嵌结构,适合其他囊泡在细胞内移动时的结合和/或解离。几种旁系同源物存在于纤毛中。在细胞皮层中发现了一组肌动蛋白,肌动蛋白勾勒出规则的表面模式。口腔特定结构的标记归因于其他类型的肌动蛋白,而更多类型则以一种表明与众多高尔基体区域相关的模式分布。相当一部分肌动蛋白与已知由其他蛋白质组成的细胞骨架成分相关。相应肌动蛋白基因或基因亚家族的沉默对与定位研究相符的细胞器产生抑制作用。敲除在分裂沟中发现的肌动蛋白会导致细胞分裂停止,而沉默其他肌动蛋白基因则会改变活力、细胞形状和游泳行为。