Chatre Laurent, Brandizzi Federica, Hocquellet Agnès, Hawes Chris, Moreau Patrick
Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, Centre National de la Recherche Scientifique-Université Bordeaux 2, 33076 Bordeaux, France.
Plant Physiol. 2005 Nov;139(3):1244-54. doi: 10.1104/pp.105.067447. Epub 2005 Oct 21.
Distinct sets of soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors (SNAREs) are distributed to specific intracellular compartments and catalyze membrane fusion events. Although the central role of these proteins in membrane fusion is established in nonplant systems, little is known about their role in the early secretory pathway of plant cells. Analysis of the Arabidopsis (Arabidopsis thaliana) genome reveals 54 genes encoding SNARE proteins, some of which are expected to be key regulators of membrane trafficking between the endoplasmic reticulum (ER) and the Golgi. To gain insights on the role of SNAREs of the early secretory pathway in plant cells, we have cloned the Arabidopsis v-SNAREs Sec22, Memb11, Bet11, and the t-SNARE Sed5, and analyzed their distribution in plant cells in vivo. By means of live cell imaging, we have determined that these SNAREs localize at the Golgi apparatus. In addition, Sec22 was also distributed at the ER. We have then focused on understanding the function of Sec22 and Memb11 in comparison to the other SNAREs. Overexpression of the v-SNAREs Sec22 and Memb11 but not of the other SNAREs induced collapse of Golgi membrane proteins into the ER, and the secretion of a soluble secretory marker was abrogated by all SNAREs. Our studies suggest that Sec22 and Memb11 are involved in anterograde protein trafficking at the ER-Golgi interface.
不同的可溶性N-乙基马来酰亚胺敏感融合蛋白附着蛋白受体(SNARE)家族分布于特定的细胞内区室,并催化膜融合事件。尽管这些蛋白在膜融合中的核心作用在非植物系统中已得到确立,但它们在植物细胞早期分泌途径中的作用却鲜为人知。对拟南芥(Arabidopsis thaliana)基因组的分析揭示了54个编码SNARE蛋白的基因,其中一些有望成为内质网(ER)与高尔基体之间膜运输的关键调节因子。为了深入了解植物细胞早期分泌途径中SNARE的作用,我们克隆了拟南芥的v-SNAREs Sec22、Memb11、Bet11以及t-SNARE Sed5,并在体内分析了它们在植物细胞中的分布。通过活细胞成像,我们确定这些SNARE定位于高尔基体。此外,Sec22也分布在内质网。然后,我们着重比较Sec22和Memb11与其他SNARE的功能。v-SNAREs Sec22和Memb11的过表达而非其他SNARE的过表达导致高尔基体膜蛋白向内质网塌陷,并且所有SNARE均抑制了可溶性分泌标记物的分泌。我们的研究表明,Sec22和Memb11参与内质网-高尔基体界面的顺向蛋白运输。