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Biogenesis of Plant Prevacuolar Multivesicular Bodies 被引量:9
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作者 Yong Cui Jinbo Shen +3 位作者 Caiji Gao Xiaohong Zhuang Junqi Wang Liwen Jiang 《Molecular Plant》 SCIE CAS CSCD 2016年第6期774-786,共13页
Plant prevacuolar compartments (PVCs), or multivesicular bodies (MVBs), are single membrane-bound organelles that play important roles in mediating protein trafficking to vacuoles in the secretory pathway. PVC/MVB... Plant prevacuolar compartments (PVCs), or multivesicular bodies (MVBs), are single membrane-bound organelles that play important roles in mediating protein trafficking to vacuoles in the secretory pathway. PVC/MVB also serves as a late endosome in the endocytic pathway in plants. Since the plant PVC was iden- tified as an MVB more than 10 years ago,-great progress has been made toward the understanding of PVC/ MVB function and biogenesis in plants. In this review, we first summarize previous research into the iden- tification and characterization of plant PVCs/MVBs, and then highlight recent advances on the mechanisms underlying intraluminal vesicle formation and maturation of plant PVCs/MVBs. In addition, we discuss the possible crosstalk that appears to occur between PVCs/MVBs and autophagosomes during autophagy in plants. Finally, we list some open questions and present future perspectives in this field. 展开更多
关键词 organelle biogenesis prevacuolar compartment multivesicular body vacuolar sorting receptor Rab5GTPase
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Retromer Subunits VPS35A and VPS29 Mediate Prevacuolar Compartment (PVC) Function in Arabidopsis
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作者 Tomasz Nodzynski Mugurel I. Ferarub +7 位作者 Sibylle Hirsch Riet De Rycke Claudiu Niculaes Wout Boerjan Jelle Van Leene Geert De Jaeger Steffen Vanneste Jiri Friml 《Molecular Plant》 SCIE CAS CSCD 2013年第6期1849-1862,共14页
Intracellular protein routing is mediated by vesicular transport which is tightly regulated in eukaryotes. The protein and lipid homeostasis depends on coordinated delivery of de novo synthesized or recycled cargoes t... Intracellular protein routing is mediated by vesicular transport which is tightly regulated in eukaryotes. The protein and lipid homeostasis depends on coordinated delivery of de novo synthesized or recycled cargoes to the plasma membrane by exocytosis and their subsequent removal by rerouting them for recycling or degradation. Here, we report the characterization of protein affected trafficking 3 (pat3) mutant that we identified by an epifluorescence-based for- ward genetic screen for mutants defective in subcellular distribution of Arabidopsis auxin transporter PIN1-GFR While pat3 displays largely normal plant morphology and development in nutrient-rich conditions, it shows strong ectopic intracellular accumulations of different plasma membrane cargoes in structures that resemble prevacuolar compart- ments (PVC) with an aberrant morphology. Genetic mapping revealed that pat3 is defective in vacuolar protein sorting 35A (VPS35A), a putative subunit of the retromer complex that mediates retrograde trafficking between the PVC and trans-Golgi network. Similarly, a mutant defective in another retromer subunit, vps29, shows comparable subcellular defects in PVC morphology and protein accumulation. Thus, our data provide evidence that the retromer components VPS35A and VPS29 are essential for normal PVC morphology and normal trafficking of plasma membrane proteins in plants. In addition, we show that, out of the three VPS35 retromer subunits present in Arabidopsis thaliana genome, the VPS35 homolog A plays a prevailing role in trafficking to the lyric vacuole, presenting another level of complexity in the retromer-dependent vacuolar sorting. 展开更多
关键词 RETROMER VPS35 VPS29 prevacuolar compartment (PVC) vacuolar trafficking Arabidopsis thaliana.
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Molecular Characterization of Plant Prevacuolar and Endosomal Compartments
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作者 Sheung Kwan Lam Yu Chung Tse +4 位作者 Yansong Miao Hong-Ye Li Junqi Wang Sze Wan Lo Liwen Jiang 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2007年第8期1119-1128,共10页
Prevacuolar compartments (PVCs) and endosomal compartments are membrane-bound organelles mediating protein traffic to vacuoles in the secretory and endocytic pathways of plant cells. Over the years, great progress h... Prevacuolar compartments (PVCs) and endosomal compartments are membrane-bound organelles mediating protein traffic to vacuoles in the secretory and endocytic pathways of plant cells. Over the years, great progress has been made towards our understanding in these two compartments in plant cells. In this review, we will summarize our contributions toward the identification and characterization of plant prevacuolar and endosomal compartments. Our studies will serve as important steps in future molecular characterization of PVC biogenesis and PVC-mediated protein traffickinq in plant cells. 展开更多
关键词 BY-2 cells endosomal compartment prevacuolar compartment organelle proteomic vacuolar sorting receptor
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水稻小G蛋白OsRab5b的亚细胞定位研究 被引量:1
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作者 邵军丽 龙跃生 徐增富 《生物技术通报》 CAS CSCD 北大核心 2015年第11期139-145,共7页
旨在研究水稻Os Rab5b亚细胞定位及第二位甘氨酸在蛋白定位中的作用。利用药物、细胞器标记物和示踪染料处理Os Rab5b-GFP与Os Rab5b(Gly2Ala)-GFP的转基因BY-2细胞,然后用激光共聚焦显微镜观察。结果表明,Os Rab5b-GFP转基因细胞呈现... 旨在研究水稻Os Rab5b亚细胞定位及第二位甘氨酸在蛋白定位中的作用。利用药物、细胞器标记物和示踪染料处理Os Rab5b-GFP与Os Rab5b(Gly2Ala)-GFP的转基因BY-2细胞,然后用激光共聚焦显微镜观察。结果表明,Os Rab5b-GFP转基因细胞呈现出大量的分散点状结构和少量细胞质弥散信号;wortmannin处理可以使Os Rab5b-GFP标记的点状结构膨胀成小的环状结构;采用100μg/m L布雷菲德菌素A(BFA)处理可使Os Rab5b-GFP标记的结构聚集。大部分的Os Rab5b-GFP信号都可以与前液泡区标记蛋白VSRAt-1共定位。在内吞示踪染料FM4-64摄取的第60 min,Os Rab5b-GFP标记的结构大部分被FM4-64标记。突变体Os Rab5b(Gly2Ala)-GFP弥散分布于细胞核和细胞质内,而且不受wortmannin或BFA药物处理的影响。Os Rab5b定位于BY-2细胞的前液泡区,Gly2在蛋白准确定位方面发挥重要作用。 展开更多
关键词 OsRab5b 亚细胞定位 前液泡区 内吞体
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从烟草悬浮细胞中分离液泡前体的方法研究 被引量:1
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作者 莫蓓莘 姜里文 《深圳大学学报(理工版)》 EI CAS 北大核心 2005年第2期109-112,共4页
通过跟踪含有液泡前体特异标记物的组分, 利用细胞壁酶水解胞壁制造原生质体, 并用真针头挤压的方法裂解细胞, 用蔗糖密度梯度离心的方法分离液泡前体. 通过相差显微镜观察, 发现所分离的液泡前体保持完整的结构. 利用不同细胞器的标记... 通过跟踪含有液泡前体特异标记物的组分, 利用细胞壁酶水解胞壁制造原生质体, 并用真针头挤压的方法裂解细胞, 用蔗糖密度梯度离心的方法分离液泡前体. 通过相差显微镜观察, 发现所分离的液泡前体保持完整的结构. 利用不同细胞器的标记物抗体进行免疫标记检测, 证明所纯化的液泡前体不含其他细胞器, 具有较高的纯度. 展开更多
关键词 前体 液泡 分离 悬浮细胞 蔗糖密度梯度离心 烟草 显微镜观察 原生质体 免疫标记 标记物 细胞器 酶水解 细胞壁
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烟草液泡前体的分离与超微结构研究
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作者 莫蓓莘 姜里文 《深圳大学学报(理工版)》 EI CAS 北大核心 2005年第3期248-252,共5页
从含有液泡前体荧光标记物的烟草转基因细胞系中分离纯化液泡前体.用荧光共聚焦显微镜跟踪观察纯化过程,明显观察到荧光信号的逐渐增强.纯化后的组分经负染后直接用电子显微镜观察,可见大小形态均一的细胞器,经脱水包埋超薄切片处理后... 从含有液泡前体荧光标记物的烟草转基因细胞系中分离纯化液泡前体.用荧光共聚焦显微镜跟踪观察纯化过程,明显观察到荧光信号的逐渐增强.纯化后的组分经负染后直接用电子显微镜观察,可见大小形态均一的细胞器,经脱水包埋超薄切片处理后在电子显微镜下可见多小泡体结构的细胞器,该细胞器可被液泡前体的特异标记物VSR标记.荧光信号的逐渐增强和液泡前体特异标记物的标记结果证明,所分离得到的多小泡体是液泡前体. 展开更多
关键词 液泡前体 荧光共聚焦显微镜 电子显微镜 烟草悬浮细胞 多小泡体
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液泡前体蛋白质的双向电泳分离和质谱分析
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作者 莫蓓莘 姜里文 《华东师范大学学报(自然科学版)》 CAS CSCD 北大核心 2005年第2期91-97,共7页
该文通过改进双向电泳方法克服了膜蛋白由于有疏水性而很难进入双向电泳第一相——等电聚焦电泳凝胶,以及由于从蔗糖密度梯度离心中分离得到的液泡前体富含干扰等电聚焦电泳效果的蔗糖等两大难点,成功地分离了通过蔗糖密度梯度离心分离... 该文通过改进双向电泳方法克服了膜蛋白由于有疏水性而很难进入双向电泳第一相——等电聚焦电泳凝胶,以及由于从蔗糖密度梯度离心中分离得到的液泡前体富含干扰等电聚焦电泳效果的蔗糖等两大难点,成功地分离了通过蔗糖密度梯度离心分离得到的液泡前体蛋白,并使膜蛋白进入了双向电泳凝胶.双向电泳后分离到约200余个蛋白质点,用基质辅助激光解吸/电离飞行时间串联质谱(MS/MS)进行肽质量指纹谱分析,并通过数据库检索进行蛋白质鉴定与功能预测,分析的23个蛋白中有13个在数据库中没有找到相对应的吻合蛋白,查询到的10个吻合蛋白中有6个功能未知. 展开更多
关键词 双向电泳 肽质量指纹谱 液泡前体 膜蛋白 烟草
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K+ Transporter AtCHX17 with Its Hydrophilic C Tail Localizes to Membranes of the Secretory/Endocytic System: Role in Reproduction and Seed Set 被引量:5
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作者 Salil Chanroj Senthilkumar Padmanaban +2 位作者 Daniel D. Czerny Guang-Yuh Jauh Heven Sze 《Molecular Plant》 SCIE CAS CSCD 2013年第4期1226-1246,共21页
The importance of sorting proteins and wall materials to their destination is critical for plant growth and development, though the machinery orchestrating membrane trafficking is poorly understood. Transporters that ... The importance of sorting proteins and wall materials to their destination is critical for plant growth and development, though the machinery orchestrating membrane trafficking is poorly understood. Transporters that alter the environment across endomembrane compartments are thought to be important players. Using Escherichia coli and yeast, we previously showed that several Arabidopsis Cation/H+ eXchanger (AtCHX) members were K+ transporters with a role in pH homeostasis, though their subcellular location and biological roles in plants are unclear. Co-expression of markers with CHX16, CHX17, CHX18, or CHX19 tagged with a fluorescent protein indicated these transporters associated with plasma membrane (PM) and post-Golgi compartments. Under its native promoter, AtCHX17(l_820)-GFP localized to prevacuolar compartment (PVC) and to PM in roots. Brefeldin A diminished AtCHX17- GFP fluorescence at PM, whereas wortmannin caused formation of GFP-labeled ring-like structures, suggesting AtCHX17 trafficked among PVC, vacuole and PM. AtCHX17(1-472) lacking its carboxylic tail did not associate with PVC or PM in plant cells. Single chx17 mutant or higher-order mutants showed normal root growth and vegetative devel- opment. However, quadruple (chx16chx17chxlSchx19) mutants were reduced in frequency and produced 50%-70% fewer seeds, indicating overlapping roles of several AtCHX17-related transporters in reproduction and/or seed devel- opment. Together, our results suggest that successful reproduction and seed development depend on the ability to regulate cation and pH homeostasis by AtCHX17-1ike transporters on membranes that traffic in the endocytic and/or secretory pathways. 展开更多
关键词 membrane trafficking pH homeostasis cation/proton antiporter prevacuolar compartment.
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