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Plant and Cell Physiology, 2003, Vol. 44, No. 8 795-802
© 2003 Oxford University Press

Alpha Tonoplast Intrinsic Protein is Specifically Associated with Vacuole Membrane Involved in an Autophagic Process

Yuji Moriyasu1, Masaki Hattori1, Guang-Yuh Jauh2,3 and John C. Rogers2,4

1 School of Food and Nutritional Sciences, University of Shizuoka, 52-1 Yada, Shizuoka, Japan
2 Institute of Biological Chemistry, Washington State University, Pullman, WA 99164-6340 U.S.A.

Autophagy in plant cells is induced by nutrient starvation. Initially, double membrane-bound organelles, termed autophagosomes, enclose a portion of cytoplasm, and then fuse with a vacuole or lysosome to give an autolysosome. Autolysosomes can be visualized by incubating cells in the presence of a membrane-permeable cysteine protease inhibitor. The inhibitor presumably decreases proteolytic degradation of the autolysosome contents that are composed of portions of cytoplasm enclosed by the membrane originating from the inner membrane of autophagosomes, and allows them to accumulate. The origin of membranes that give rise to autophagosomes and autolysosomes is unknown. Here we use an acidotropic fluorescent dye, LysoTracker Red, to label autolysosomes specifically. We demonstrate that autolysosome membranes are marked by the presence of {alpha}-tonoplast intrinsic protein ({alpha}-TIP) but not by {gamma}-TIP or {delta}-TIP. The identification of a TIP specifically associated with membranes derived from an autophagic process may help our understanding of how plant cells generate and maintain functionally distinct types of vacuoles.

4 Corresponding author: E-mail: bcjroger{at}wsu.edu; Fax, +1-509-335-7643.

3 Present address: Institute of Botany, Academia Sinica, Nankang, Taipei 11529, Taiwan


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