Department of Biology, Centre for Cell and Developmental Biology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
Plant J. 2010 Mar;61(5):826-38. doi: 10.1111/j.1365-313X.2009.04111.x. Epub 2009 Dec 15.
Vacuolar sorting receptors (VSRs) are type-I integral membrane proteins that mediate biosynthetic protein traffic in the secretory pathway to the vacuole, whereas secretory carrier membrane proteins (SCAMPs) are type-IV membrane proteins localizing to the plasma membrane and early endosome (EE) or trans-Golgi network (TGN) in the plant endocytic pathway. As pollen tube growth is an extremely polarized and highly dynamic process, with intense anterograde and retrograde membrane trafficking, we have studied the dynamics and functional roles of VSR and SCAMP in pollen tube growth using lily (Lilium longiflorum) pollen as a model. Using newly cloned lily VSR and SCAMP cDNA (termed LIVSR and LISCAMP, respectively), as well as specific antibodies against VSR and SCAMP1 as tools, we have demonstrated that in growing lily pollen tubes: (i) transiently expressed GFP-VSR/GFP-LIVSR is located throughout the pollen tubes, excepting the apical clear-zone region, whereas GFP-LISCAMP is mainly concentrated in the tip region; (ii) VSRs are localized to the multivesicular body (MVB) and vacuole, whereas SCAMPs are localized to apical endocytic vesicles, TGN and vacuole; and (iii) microinjection of VSR or SCAMP antibodies and LlVSR small interfering RNAs (siRNAs) significantly reduced the growth rate of the lily pollen tubes. Taken together, both VSR and SCAMP are required for pollen tube growth, probably working together in regulating protein trafficking in the secretory and endocytic pathways, which need to be coordinated in order to support pollen tube elongation.
液泡分选受体(VSRs)是一类Ⅰ型整合膜蛋白,它们介导生物合成蛋白在分泌途径中向液泡的运输,而分泌载体膜蛋白(SCAMPs)则是定位于植物内吞途径中的质膜和早期内体(EE)或反式高尔基体网络(TGN)的Ⅳ型膜蛋白。由于花粉管的生长是一个极度极化和高度动态的过程,具有强烈的正向和逆向膜运输,我们使用百合(Lilium longiflorum)花粉作为模型,研究了 VSR 和 SCAMP 在花粉管生长中的动态和功能作用。使用新克隆的百合 VSR 和 SCAMP cDNA(分别称为 LIVSR 和 LISCAMP)以及针对 VSR 和 SCAMP1 的特异性抗体作为工具,我们证明了在生长的百合花粉管中:(i)瞬时表达的 GFP-VSR/GFP-LIVSR 遍布花粉管,除了顶端透明区区域,而 GFP-LISCAMP 主要集中在尖端区域;(ii)VSR 定位于多泡体(MVB)和液泡,而 SCAMPs 定位于顶端内吞小泡、TGN 和液泡;(iii)VSR 或 SCAMP 抗体和 LlVSR 小干扰 RNA(siRNA)的微注射显著降低了百合花粉管的生长速度。总之,VSR 和 SCAMP 都对花粉管的生长是必需的,可能共同作用于调节分泌和内吞途径中的蛋白质运输,需要协调以支持花粉管伸长。