Ebine Kazuo, Okatani Yusuke, Uemura Tomohiro, Goh Tatsuaki, Shoda Keiko, Niihama Mitsuru, Morita Miyo Terao, Spitzer Christoph, Otegui Marisa S, Nakano Akihiko, Ueda Takashi
Department of Biological Sciences, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
Plant Cell. 2008 Nov;20(11):3006-21. doi: 10.1105/tpc.107.057711. Epub 2008 Nov 4.
The SNARE complex is a key regulator of vesicular traffic, executing membrane fusion between transport vesicles or organelles and target membranes. A functional SNARE complex consists of four coiled-coil helical bundles, three of which are supplied by Q-SNAREs and another from an R-SNARE. Arabidopsis thaliana VAMP727 is an R-SNARE, with homologs only in seed plants. We have found that VAMP727 colocalizes with SYP22/ VAM3, a Q-SNARE, on a subpopulation of prevacuolar compartments/endosomes closely associated with the vacuolar membrane. Genetic and biochemical analyses, including examination of a synergistic interaction of vamp727 and syp22 mutations, histological examination of protein localization, and coimmunoprecipitation from Arabidopsis lysates indicate that VAMP727 forms a complex with SYP22, VTI11, and SYP51 and that this complex plays a crucial role in vacuolar transport, seed maturation, and vacuole biogenesis. We suggest that the VAMP727 complex mediates the membrane fusion between the prevacuolar compartment and the vacuole and that this process has evolved as an essential step for seed development.
SNARE复合体是囊泡运输的关键调节因子,负责执行运输囊泡或细胞器与靶膜之间的膜融合。一个功能性的SNARE复合体由四个卷曲螺旋螺旋束组成,其中三个由Q-SNARE提供,另一个由R-SNARE提供。拟南芥VAMP727是一种R-SNARE,仅在种子植物中有同源物。我们发现VAMP727与Q-SNARE SYP22/VAM3在与液泡膜紧密相关的前液泡区室/内体的一个亚群上共定位。遗传和生化分析,包括对vamp727和syp22突变的协同相互作用的检测、蛋白质定位的组织学检查以及从拟南芥裂解物中进行的免疫共沉淀,表明VAMP727与SYP22、VTI11和SYP51形成复合体,并且该复合体在液泡运输、种子成熟和液泡生物发生中起关键作用。我们认为VAMP727复合体介导前液泡区室与液泡之间的膜融合,并且这一过程已演变为种子发育的一个重要步骤。