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THE PLANT CELL, Vol 5, Issue 9 1113-1124, Copyright © 1993 by American Society of Plant Biologists
Tonoplast and Soluble Vacuolar Proteins Are Targeted by Different Mechanisms
L. Gomez and M. J. Chrispeels
Department of Biology, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093-0116
The delivery of proteins to the vacuole and its limiting membrane (the
tonoplast) by the secretory system is thought to be a dissociative process
in which vesicles bud from one compartment and fuse with another. We
studied the transport kinetics of phytohemagglutinin (PHA) and tonoplast
intrinsic protein (TIP) in mesophyll protoplasts obtained from transgenic
tobacco plants transformed with genes encoding these two proteins. In
pulse-chase experiments, arrival of PHA in the vacuole was found to be
slower (completed 24 hr after synthesis) than the arrival of TIP in the
tonoplast (completed 6 hr after synthesis). Brefeldin A and monensin block
protein transport by interfering in specific vesicle transport steps.
Brefeldin A prevents anterograde vesicle transport between the endoplasmic
reticulum and the Golgi, whereas monensin inhibits correct sorting in the
trans-Golgi network by disrupting the proton gradient across the membrane.
Both inhibitors blocked the transport of PHA to the vacuole and altered the
rate at which its complex glycan is processed by Golgi enzymes. Neither
drug stopped the arrival of TIP in the tonoplast, suggesting that the flow
of vesicles continues in the presence of these inhibitors. We suggest that
soluble proteins like PHA and membrane proteins like TIP reach their
vacuolar destinations by different paths.
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