Zhao H, Nonet M L
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Mol Biol Cell. 2001 Aug;12(8):2275-89. doi: 10.1091/mbc.12.8.2275.
We have studied the localization of synaptogyrin family members in vivo. Both native and green fluorescent protein (GFP)-tagged Caenorhabditis elegans synaptogyrin (SNG-1) are expressed in neurons and synaptically localized. Deletion and mutational analysis with the use of GFP-tagged SNG-1 has defined a 38 amino acid sequence within the C terminus of SNG-1 and a single arginine in the cytoplasmic loop between transmembrane domain 2 and 3 that are required for SNG-1 localization. These domains may represent components of signals that target synaptogyrin for endocytosis from the plasma membrane and direct synaptogyrin to synaptic vesicles, respectively. In chimeric studies, these regions were sufficient to relocalize cellugyrin, a nonneuronal form of synaptogyrin, from nonsynaptic regions such as the sensory dendrites and the cell body to synaptic vesicles. Furthermore, GFP-tagged rat synaptogyrin is synaptically localized in neurons of C. elegans and in cultured hippocampal neurons. Similarly, the C-terminal domain of rat synaptogyrin is necessary for localization in hippocampal neurons. Our study suggests that the mechanisms for synaptogyrin localization are likely to be conserved from C. elegans to vertebrates.
我们已经在体内研究了突触素家族成员的定位。线虫天然的和绿色荧光蛋白(GFP)标记的突触素(SNG-1)均在神经元中表达并定位于突触。利用GFP标记的SNG-1进行的缺失和突变分析确定了SNG-1 C末端的一个38个氨基酸的序列以及跨膜结构域2和3之间细胞质环中的一个精氨酸,它们是SNG-1定位所必需的。这些结构域可能分别代表将突触素从质膜靶向内吞以及将突触素导向突触小泡的信号成分。在嵌合研究中,这些区域足以将细胞突触素(一种非神经元形式的突触素)从诸如感觉树突和细胞体等非突触区域重新定位到突触小泡。此外,GFP标记的大鼠突触素定位于线虫神经元和培养的海马神经元的突触中。同样,大鼠突触素的C末端结构域对于在海马神经元中的定位是必需的。我们的研究表明,从线虫到脊椎动物,突触素定位机制可能是保守的。