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Analysis of Endocytosis and Ubiquitination of the BOR1 Transporter

  • Koji KasaiEmail author
  • Junpei Takano
  • Toru Fujiwara
Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1209)

Abstract

Endocytosis and membrane trafficking are the major factors controlling the abundance of plasma membrane proteins, such as transporters and receptors. We have found that Arabidopsis borate transporter BOR1 is polarly localized to the inner (stele-facing) plasma membrane domain of various root cells under boron limitation, and when boron is supplied in excess, BOR1 is rapidly transferred to the vacuole for immediate degradation. The BOR1 polarity and degradation are controlled by membrane trafficking including endocytosis. In this chapter, we describe methods for observation of endocytic trafficking of BOR1, and detection of BOR1 ubiquitination that is required for vacuolar sorting for degradation.

Key words

Endocytosis Membrane trafficking BOR1 Polar localization Boron-induced degradation Vacuolar sorting Ubiquitination 
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Notes

Acknowledgement

This work was supported by a Grant-in-Aid for JSPS Fellows (no. 19-7094 to K.K.) from the Japan Society for the Promotion of Science, by a Grant-in-Aid for Young Scientists (Wakate B-237800060, to K.K.) from the Ministry of Education, Culture, Sports, Science and Technology, Japan, by a Grant-in-Aid for Scientific Research (to T.F.), and a Grant-in-Aid for Scientific Research Priority Areas (to T.F.) from the Ministry of Education, Culture, Sports, Science and Technology, Japan, by the NEXT program (to J.T.) from the Japanese Society for the Promotion of Science.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Graduate School of Agricultural and Life SciencesThe University of TokyoTokyoJapan
  2. 2.Research Faculty of AgricultureHokkaido UniversitySapporoJapan

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