Plant Physiology Preview
Published on October 13, 2006; 10.1104/pp.106.090423
OPEN ACCESS ARTICLE
Received September 27, 2006
Accepted October 9, 2006
Dynamic Response of Prevacuolar Compartments to Brefeldin A in Plant Cells
Yu Chung Tse , Sze Wan Lo , Stefan Hillmer , Paul Dupree , and Liwen Jiang *
Department of Biology; Molecular Biotechnology Program, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China
Department of Cell Biology, Heidelberg Institute for Plant Sciences, University of Heidelberg, D-69120 Heidelberg, Germany
Department of Biochemistry, University of Cambridge, UK
* Corresponding author; email: ljiang{at}cuhk.edu.hk.
Little is known about the dynamics and molecular components of plant prevacuolar compartments (PVCs) in the secretory pathway. Using transgenic tobacco BY-2 cells expressing membrane-anchored yellow fluorescent protein (YFP) reporters marking Golgi or PVCs, we have recently demonstrated that PVCs are mobile multivesicular bodies (MVBs) defined by vacuolar sorting receptor (VSR) proteins. Here we demonstrate that Golgi and PVCs have different sensitivity in response to Brefeldin A (BFA) treatment in living tobacco BY-2 cells. BFA at low concentrations (5 to 10 µg mL-1) induced YFP-marked Golgi stacks to form both ER-Golgi hybrid structures and BFA-induced aggregates, but had little effect on YFP-marked PVCs in transgenic BY-2 cells at both confocal and immunoEM levels. However, BFA at high concentrations (50 to 100 µg mL-1) caused both YFP-marked Golgi stacks and PVCs to form aggregates in a dosage-dependent and time-dependent manner. Normal Golgi or PVC signals can be recovered upon removal of BFA from the culture media. Confocal immunofluorescence and immunoEM studies with specific organelle markers further demonstrate that the PVC aggregates are distinct but physically associated with Golgi aggregates in BFA-treated cells and that PVCs might lose their internal vesicle structures at high BFA concentration. In addition, VSR-marked PVCs in root tip cells of tobacco, pea, mungbean and Arabidopsis upon BFA treatment are also induced to form similar aggregates. Thus, we have demonstrated that BFA's effects are not limited to ER and Golgi but extended to PVC in the endomembrane system, which might provide a quick tool for distinguishing Golgi from PVC for its identification and characterization, as well as a possible new tool in studying PVC-mediated protein traffic in plant cells.
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