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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Divergent functions of VTI12 and VTI11 in trafficking to storage and Iytic vacuoles in Arabidopsis
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Divergent functions of VTI12 and VTI11 in trafficking to storage and Iytic vacuoles in Arabidopsis

机译：VTI12和VTI11在拟南芥中运输到储存和Iytic空泡的功能不同

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The protein storage vacuole (PSV) is a plant-specific organelle that accumulates reserve proteins, one of the main agricultural products obtained from crops. Despite the importance of this process, the cellular machinery required for transport and accumulation of storage proteins remains largely unknown. Interfering with transport to PSVs has been shown to result in secretion of cargo. Therefore, secretion of a suitable marker could be used as an assay to identify mutants in this pathway. CLV3, a negative regulator of shoot stem cell proliferation, is an extracellular ligand that is rendered inactive when targeted to vacuoles. We devised an assay where trafficking mutants secrete engineered vacuolar CLV3 and show reduced meristems, a phenotype easily detected by visual inspection of plants. We tested this scheme in plants expressing VAC2, a fusion of CLV3 to the vacuolar sorting signal from the storage protein barley lectin. In this way, we determined that trafficking of VAC2 requires the SNARE VTI12 but not its close homologue, the conditionally redundant VTI11 protein. Furthermore, a vti12 mutant is specifically altered in transport of storage proteins, whereas a vti11 mutant is affected in transport of a lytic vacuole marker. These results demonstrate the specialization of VTI12 and VTI11 in mediating trafficking to storage and lytic vacuoles, respectively. Moreover, they validate the VAC2 secretion assay as a simple method to isolate genes that mediate trafficking to the PSV.
机译：蛋白质储存液泡（PSV）是一种植物特有的细胞器，可累积储备蛋白质，而储备蛋白质是从农作物中获得的主要农产品之一。尽管此过程很重要，但运输和积累存储蛋白所需的细胞机制仍然未知。已证明干扰到PSV的运输会导致货物分泌。因此，合适标记的分泌可以用作鉴定该途径中的突变体的测定法。 CLV3是芽干细胞增殖的负调节剂，是一种细胞外配体，当靶向液泡时会失活。我们设计了一种分析方法，其中贩运突变体分泌工程化的液泡CLV3并显示出分生组织减少，这种分型易于通过肉眼检查植物来检测。我们在表达VAC2的植物中测试了该方案，VAC2是CLV3与来自存储蛋白大麦凝集素的液泡分选信号的融合体。通过这种方式，我们确定VAC2的运输需要SNARE VTI12，而不是其紧密同源物，即条件冗余的VTI11蛋白。此外，vti12突变体在存储蛋白的运输中发生了特定变化，而vti11突变体在溶性液泡标记物的运输中受到影响。这些结果证明了VTI12和VTI11分别在介导运输到储存和溶解液泡中的专长。此外，他们验证了VAC2分泌测定法是一种简单的方法，可将介导贩运至PSV的基因分离出来。

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