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Golgi Apparatus-localized Synaptotagmin 2 is Required for Unconventional Trafficking/Secretory Pathway in Arabidopsis

作　　者：Haiyan Zhang, Liang Zhang, Bin Gao, Hai Fan, Jingbo Jin, Miguel A. Botella, Liwen Jiang, Jinxing Lin
影响因子：4.411
刊物名称：PLoS ONE
出版年份：2011
卷：6  期：11  页码：e26477

论文摘要：

 Background: Most secretory proteins contain signal peptides that direct their sorting to the ER and secreted via the conventional ER/Golgi transport pathway, while some signal peptide-lacking proteins have been shown to export through ER/Golgi independent secretory pathways. Hygromycin B is an aminoglycoside antibiotic produced by Streptomyces hygroscopicus that is active against both prokaryotic and eukaryotic cells. The hygromycin phosphotransferase (HYGR) can phosphorylate and inactivate the hygromycin B, and has been widely used as a positive selective marker in the construction of transgenic plants. However, the localization and trafficking of HYGR in plant cells remain unknown. Synaptotagmins (SYTs) are involved in controlling vesicle endocytosis and exocytosis as calcium sensors in animal cells, while their functions in plant cells are largely unclear.
Methodology/Principal Findings: We found Arabidopsis synaptotagmin SYT2 was localized on the Golgi apparatus by immunofluorescence and immunogold labeling. Surprisingly, co-expression of SYT2 and HYGR caused hypersensitivity of the transgenic Arabidopsis plants to hygromycin B. HYGR, which lacks a signal sequence, was present in the cytoplasm as well as in the extracellular space in HYGR-GFP transgenic Arabidopsis plants and its secretion is not sensitive to brefeldin A treatment, suggesting it is not secreted via the conventional secretory pathway. Furthermore, we found that HYGR-GFP was truncated at carboxyl terminus of HYGR shortly after its synthesis, and the cells deficient SYT2 failed to efficiently truncate HYGR-GFP,resulting in HYGR-GFP accumulated in prevacuoles/vacuoles, indicating that SYT2 was involved in HYGR-GFP trafficking and secretion.
Conclusion/Significance: These findings reveal for the first time that SYT2 is localized on the Golgi apparatus and regulates
HYGR-GFP secretion via the unconventional protein transport from the cytosol to the extracelluar matrix in plant cells.
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