Plant Physiology Preview
Published on November 19, 2004; 10.1104/pp.104.049411
Received July 11, 2004
Returned for revision October 6, 2004
Accepted October 12, 2004
A Green Fluorescent Protein Fusion to Actin-Binding Domain 2 of Arabidopsis Fimbrin Highlights New Features of a Dynamic Actin Cytoskeleton in Live Plant Cells
Michael B. Sheahan , Chris J. Staiger , Ray J. Rose , and David W. McCurdy *
School of Environmental and Life Sciences, The University of Newcastle, Callaghan, New South Wales, 2308 Australia; Australian Research Council Centre of Excellence for Integrative Legume Research, The University of Newcastle, Callaghan, New South Wales, 2308 Australia
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-1392
School of Environmental and Life Sciences, The University of Newcastle, Callaghan, New South Wales, 2308 Australia
* Corresponding author; email: david.mccurdy{at}newcastle.edu.au.
The actin cytoskeleton coordinates numerous cellular processes required for plant development. The functions of this network are intricately linked to its dynamic arrangement, and thus progress in understanding how actin orchestrates cellular processes relies on critical evaluation of actin organization and turnover. To investigate the dynamic nature of the actin cytoskeleton, we used a fusion protein between green fluorescent protein (GFP) and the second actin-binding domain (fABD2) of Arabidopsis (Arabidopsis thaliana) fimbrin, AtFIM1. The GFP-fABD2 fusion protein labeled highly dynamic and dense actin networks in diverse species and cell types, revealing structural detail not seen with alternative labeling methods, such as the commonly used mouse talin GFP fusion (GFP-mTalin). Further, we show that expression of the GFP-fABD2 fusion protein in Arabidopsis, unlike GFP-mTalin, has no detectable adverse effects on plant morphology or development. Time-lapse confocal microscopy and fluorescence recovery after photobleaching analyses of the actin cytoskeleton labeled with GFP-fABD2 revealed that lateral-filament migration and sliding of individual actin filaments or bundles are processes that contribute to the dynamic and continually reorganizing nature of the actin scaffold. These new observations of the dynamic actin cytoskeleton in plant cells using GFP-fABD2 reveal the value of this probe for future investigations of how actin filaments coordinate cellular processes required for plant development.
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