Polarized localization and borate-dependent degradation of the Arabidopsis borate transporter BOR1 in tobacco BY-2 cells.
In Arabidopsis the borate transporter BOR1, which is located in the plasma membrane, is degraded in the presence of excess boron by an endocytosis-mediated mechanism. A similar mechanism was suggested in rice as excess boron decreased rice borate transporter levels, although in this case whether the decrease was dependent on an increase in degradation or a decrease in protein synthesis was not elucidated. To address whether the borate-dependent degradation mechanism is conserved among plant cells, we analyzed the fate of GFP-tagged BOR1 (BOR1-GFP) in transformed tobacco BY-2 cells. Cells expressing BOR1-GFP displayed GFP fluorescence at the plasma membrane, especially at the membrane between two attached cells. The plasma membrane signal was abolished when cells were incubated in medium with a high concentration of borate (3 to 5 mM). This decrease in BOR1-GFP signal was mediated by a specific degradation of the protein after internalization by endocytosis from the plasma membrane. Pharmacological analysis indicated that the decrease in BOR1-GFP largely depends on the increase in degradation rate and that the degradation was mediated by a tyrosine-motif and the actin cytoskeleton. Tyr mutants of BOR1-GFP, which has been shown to inhibit borate-dependent degradation in Arabidopsis root cells, did not show borate-dependent endocytosis in tobacco BY-2 cells. These findings indicate that the borate-dependent degradation machinery of the borate transporter is conserved among plant species.
Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Fukuoka, 812-8581, Japan.
Noboru
N
Yamauchi
Biotechnology Research Center, The University of Tokyo, Tokyo, 113-8657, Japan.
Tadashi
T
Gosho
Laboratory of Plant Nutrition, Faculty of Agriculutre, Kyushu University, Fukuoka, 812-8581, Japan ; Current address: Omu Milk Products Co., Ltd., Omuta, 836-0895, Japan.
Satoru
S
Asatuma
RIKEN Plant Science Center, Yokohama, 230-0045, Japan ; Current address: RIKEN Center for Sustainable Resource Science, Yokohama, 230-0045, Japan.
Kiminori
K
Toyooka
Biotechnology Research Center, The University of Tokyo, Tokyo, 113-8657, Japan ; Current address: Laboratory of Plant Nutrition and Fertilizer, Graduate School of Agricultural and Life Science, The University of Tokyo, Tokyo, 113-8657, Japan.
Toru
T
Fujiwara
Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Fukuoka, 812-8581, Japan ; Laboratory of Plant Nutrition, Faculty of Agriculutre, Kyushu University, Fukuoka, 812-8581, Japan ; RIKEN Plant Science Center, Yokohama, 230-0045, Japan ; Organelle Homeostasis Research Center, Kyushu University, Fukuoka, 812-8581, Japan ; Biotron Application Center, Kyushu University, Fukuoka, 812-8581, Japan.
Ken
K
Matsuoka