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Plant Cell, Vol. 13, 287-301, February 2001, Copyright © 2001, American Society of Plant Physiologists

Trafficking of Phosphatidylinositol 3-Phosphate from the trans-Golgi Network to the Lumen of the Central Vacuole in Plant Cells

Dae Heon Kima, Young-Jae Eub, Cheol Min Yoob, Yong-Woo Kimb, Kyeong Tae Pihb, Jing Bo Jina, Soo Jin Kimb, Harald Stenmarkc, and Inhwan Hwangb
a Department of Molecular Biology, Gyeongsang National University, Chinju, 660-701, Korea
b Division of Molecular and Life Science and Center for Plant Intracellular Trafficking, Pohang University of Science and Technology, Pohang, 790-784, Korea
c Department of Biochemistry, Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway

Correspondence to: Inhwan Hwang, ihhwang{at}postech.ac.kr (E-mail), 82-562-279-8159 (fax)

Very limited information is available on the role of phosphatidylinositol 3-phosphate (PI[3]P) in vesicle trafficking in plant cells. To investigate the role of PI(3)P during the vesicle trafficking in plant cells, we exploited the PI(3)P-specific binding property of the endosome binding domain (EBD) (amino acids 1257 to 1411) of human early endosome antigen 1, which is involved in endosome fusion. When expressed transiently in Arabidopsis protoplasts, a green fluorescent protein (GFP):EBD fusion protein exhibited PI(3)P-dependent localization to various compartments—such as the trans-Golgi network, the prevacuolar compartment, the tonoplasts, and the vesicles in the vacuolar lumen—that varied with time. The internalized GFP:EBD eventually disappeared from the lumen. Deletion experiments revealed that the PI(3)P-dependent localization required the Rab5 binding motif in addition to the zinc finger motif. Overexpression of GFP:EBD inhibited vacuolar trafficking of sporamin but not trafficking of H+-ATPase to the plasma membrane. On the basis of these results, we propose that the trafficking of GFP:EBD reflects that of PI(3)P and that PI(3)P synthesized at the trans-Golgi network is transported to the vacuole through the prevacuolar compartment for degradation in plant cells.




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