A GFP-mouse talin fusion protein labels plant actin filamentsin vivoand visualizes the actin cytoskeleton in growing pollen tubes
Article first published online: 2 OCT 2008
The Plant Journal
Volume 16, Issue 3, pages 393–401, November 1998
How to Cite
Kost, B., Spielhofer, P. and Chua, N.-H. (1998), A GFP-mouse talin fusion protein labels plant actin filamentsin vivoand visualizes the actin cytoskeleton in growing pollen tubes. The Plant Journal, 16: 393–401. doi: 10.1046/j.1365-313x.1998.00304.x
- Issue published online: 2 OCT 2008
- Article first published online: 2 OCT 2008
- Received 24 August 1998; accepted 22 September 1998.
The C-terminus of mouse talin (amino acids 2345–2541) is responsible for all of the protein’s f-actin binding capacity. Unlike full-length talin, the C-terminal f-actin binding domain is unable to nucleate actin polymerization. We have found that transient and stable expression of the talin actin-binding domain fused to the C-terminus of the green fluorescent protein (GFP-mTn) can visualize the actin cytoskeleton in different types of living plant cells without affecting cell morphology or function. Transiently expressed GFP-mTn co-localized with rhodamine-phalloidin in permeabilized tobacco BY-2 suspension cells, showing that the fusion protein can specifically label the plant actin cytoskeleton. Constitutive expression of GFP-mTn in transgenicArabidopsis thalianaplants visualized actin filaments in all examined tissues with no apparent effects on plant morphology or development at any stage during the life cycle. This demonstrates that in a number of different cell types GFP-mTn can serve as a non-invasive marker for the actin cytoskeleton. Confocal imaging of GFP-mTn labeled actin filaments was employed to reveal novel information on thein vivoorganization of the actin cytoskeleton in transiently transformed, normally elongating tobacco pollen tubes.