Yokoyama R, Nishitani K
Biological Institute, Graduate School of Science, Tohoku University, Sendai, 980-8578 Japan.
Plant Cell Physiol. 2001 Mar;42(3):292-300. doi: 10.1093/pcp/pce034.
Intracellular trafficking of enzymes responsible for constructing and modifying the cell wall architecture in plants is mostly unknown. To examine their translocation pathways, we employed an endoxyloglucan transferase (EXGT), a key enzyme responsible for forming and rearranging the cellulose/xyloglucan network of the cell wall. We traced its intracellular localization in suspension-cultured cells of tobacco bright yellow-2 by means of green fluorescent protein-fusion gene procedures as well as by indirect immunofluorescence. During interphase the protein was extensively secreted into the apoplast via the endoplasmic reticulum-Golgi apparatus network, whereas during cytokinesis, the protein was exclusively located in the phragmoplast and eventually transported to the cell plate. These results clearly indicate commitment of EXGT protein to the construction of both the cell plate and the cell wall. This study also visualized the process of phragmoplast development at a level of vesicle translocation in the living cell.
植物中负责构建和修饰细胞壁结构的酶的细胞内运输过程大多尚不明确。为了研究它们的转运途径,我们选用了一种内切木葡聚糖转移酶(EXGT),这是一种负责形成和重排细胞壁中纤维素/木葡聚糖网络的关键酶。我们通过绿色荧光蛋白融合基因方法以及间接免疫荧光法,追踪了它在烟草亮黄-2悬浮培养细胞中的细胞内定位。在间期,该蛋白通过内质网-高尔基体网络大量分泌到质外体中,而在胞质分裂期间,该蛋白仅位于成膜体中,并最终运输到细胞板。这些结果清楚地表明EXGT蛋白参与了细胞板和细胞壁的构建。这项研究还在活细胞中囊泡转运水平上直观呈现了成膜体的发育过程。