Lauber M H, Waizenegger I, Steinmann T, Schwarz H, Mayer U, Hwang I, Lukowitz W, Jürgens G
Lehrstuhl für Entwicklungsgenetik, Universität Tübingen, D-72076 Tübingen, Federal Republic of Germany.
J Cell Biol. 1997 Dec 15;139(6):1485-93. doi: 10.1083/jcb.139.6.1485.
In higher plant cytokinesis, plasma membrane and cell wall originate by vesicle fusion in the plane of cell division. The Arabidopsis KNOLLE gene, which is required for cytokinesis, encodes a protein related to vesicle-docking syntaxins. We have raised specific rabbit antiserum against purified recombinant KNOLLE protein to show biochemically and by immunoelectron microscopy that KNOLLE protein is membrane associated. Using immunofluorescence microscopy, KNOLLE protein was found to be specifically expressed during mitosis and, unlike the plasma membrane H+-ATPase, to localize to the plane of division during cytokinesis. Arabidopsis dynamin-like protein ADL1 accumulates at the plane of cell plate formation in knolle mutant cells as in wild-type cells, suggesting that cytokinetic vesicle traffic is not affected. Furthermore, electron microscopic analysis indicates that vesicle fusion is impaired. KNOLLE protein was detected in mitotically dividing cells of various parts of the developing plant, including seedling root, inflorescence meristem, floral meristems and ovules, and the cellularizing endosperm, but not during cytokinesis after the male second meiotic division. Thus, KNOLLE is the first syntaxin-like protein that appears to be involved specifically in cytokinetic vesicle fusion.
在高等植物胞质分裂过程中,质膜和细胞壁通过细胞分裂平面上的囊泡融合形成。拟南芥的KNOLLE基因是胞质分裂所必需的,它编码一种与囊泡对接SNARE蛋白相关的蛋白质。我们制备了针对纯化重组KNOLLE蛋白的特异性兔抗血清,以通过生化方法和免疫电子显微镜证明KNOLLE蛋白与膜相关。利用免疫荧光显微镜观察发现,KNOLLE蛋白在有丝分裂期间特异性表达,并且与质膜H⁺-ATP酶不同,在胞质分裂期间定位于分裂平面。拟南芥动力蛋白样蛋白ADL1在knolle突变体细胞中与野生型细胞一样,在细胞板形成平面上积累,这表明胞质分裂囊泡运输不受影响。此外,电子显微镜分析表明囊泡融合受损。在发育中植物的各个部位有丝分裂的细胞中都检测到了KNOLLE蛋白,包括幼苗根、花序分生组织、花分生组织和胚珠,以及正在细胞化的胚乳,但在雄配子第二次减数分裂后的胞质分裂过程中未检测到。因此,KNOLLE是第一个似乎专门参与胞质分裂囊泡融合的类SNARE蛋白。