Present address: Department of Parasitology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
Dynamic behavior of clathrin in Arabidopsis thaliana unveiled by live imaging
Article first published online: 14 OCT 2011
© 2011 The Authors. The Plant Journal © 2011 Blackwell Publishing Ltd
The Plant Journal
Volume 69, Issue 2, pages 204–216, January 2012
How to Cite
Ito, E., Fujimoto, M., Ebine, K., Uemura, T., Ueda, T. and Nakano, A. (2012), Dynamic behavior of clathrin in Arabidopsis thaliana unveiled by live imaging. The Plant Journal, 69: 204–216. doi: 10.1111/j.1365-313X.2011.04782.x
- Issue published online: 6 JAN 2012
- Article first published online: 14 OCT 2011
- Accepted manuscript online: 12 SEP 2011 03:45PM EST
- Received 8 August 2011; revised 5 September 2011; accepted 7 September 2011; published online 14 October 2011.
- Arabidopsis thaliana;
- live imaging;
- cell plate;
- plasma membrane;
- brefeldin A;
Clathrin-coated vesicles (CCV) are necessary for selective transport events, including receptor-mediated endocytosis on the plasma membrane and cargo molecule sorting in the trans-Golgi network (TGN). Components involved in CCV formation include clathrin heavy and light chains and several adaptor proteins that are conserved among plants. Clathrin-dependent endocytosis has been shown to play an integral part in plant endocytosis. However, little information is known about clathrin dynamics in living plant cells. In this study, we have visualized clathrin in Arabidopsis thaliana by tagging clathrin light chain with green fluorescent protein (CLC–GFP). Quantitative evaluations of colocalization demonstrate that the majority of CLC–GFP is localized to the TGN, and a minor population is associated with multivesicular endosomes and the Golgi trans-cisternae. Live imaging further demonstrated the presence of highly dynamic clathrin-positive tubules and vesicles, which appeared to mediate interactions between the TGNs. CLC–GFP is also targeted to cell plates and the plasma membrane. Although CLC–GFP colocalizes with a dynamin isoform at the plasma membrane, these proteins exhibit distinct distributions at newly forming cell plates. This finding indicates independent functions of CLC (clathrin light chains) and dynamin during the formation of cell plates. We have also found that brefeldin A and wortmannin treatment causes distinctly different alterations in the dynamics and distribution of clathrin-coated domains at the plasma membrane. This could account for the different effects of these drugs on plant endocytosis.