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THE PLANT CELL, Vol 9, Issue 4 533-546, Copyright © 1997 by American Society of Plant Biologists
A Vacuolar-Type H+-ATPase in a Nonvacuolar Organelle Is Required for the Sorting of Soluble Vacuolar Protein Precursors in Tobacco Cells
K. Matsuoka, T. Higuchi, M. Maeshima and K. Nakamura
Laboratory of Biochemistry, School of Agricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-01, Japan
In plant cells, vacuolar matrix proteins are separated from the secretory
proteins at the Golgi complex for transport to the vacuoles. To investigate
the involvement of vacuolar-type ATPase (V-ATPase) in the vacuolar
targeting of soluble proteins, we analyzed the effects of bafilomycin A1
and concanamycin A on the transport of vacuolar protein precursors in
tobacco cells. Low concentrations of these inhibitors caused the missorting
of several vacuolar protein precursors; sorting was more sensitive to
concanamycin A than to bafilomycin A1. Secretion of soluble proteins from
tobacco cells was also inhibited by bafilomycin A1 and concanamycin A. We
next analyzed the subcellular localization of V-ATPase. V-ATPase was found
in a wide variety of endomembrane organelles. Both ATPase activity and
ATP-dependent proton-pumping activity in the Golgi-enriched fraction were
more sensitive to concanamycin A than to bafilomycin A1, whereas these
activities in the tonoplast fraction were almost equally sensitive to both
reagents. Our observations indicate that the V-ATPase in the organelle that
was recovered in the Golgi-enriched fraction is required for the transport
of vacuolar protein precursors and that this V-ATPase is distinguishable
from the tonoplast-associated V-ATPase.
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