First published online November 19, 2004; 10.1104/pp.104.049411
Plant Physiology 136:3968-3978 (2004)
© 2004 American Society of Plant Biologists
A Green Fluorescent Protein Fusion to Actin-Binding Domain 2 of Arabidopsis Fimbrin Highlights New Features of a Dynamic Actin Cytoskeleton in Live Plant Cells1,[w]
Michael B. Sheahan,
Chris J. Staiger,
Ray J. Rose and
David W. McCurdy*
School of Environmental and Life Sciences (M.B.S., R.J.R., D.W.M.), and Australian Research Council Centre of Excellence for Integrative Legume Research (M.B.S., R.J.R.), The University of Newcastle, Callaghan, New South Wales, 2308 Australia; and Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907–1392 (C.J.S.)
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.
1 This work was supported by an Australian Research Council Centre of Excellence Grant to The University of Newcastle Node of the Centre of Excellence for Integrative Legume Research (to R.J.R.), and by the U.S. National Science Foundation (grant no. 0130576–MCB to C.J.S. and D.W.M.).
[w] The online version of this article contains Web-only data.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.049411.
* Corresponding author; e-mail david.mccurdy{at}newcastle.edu.au; fax 61–2–49–21–6923.
Received July 11, 2004;
returned for revision October 6, 2004;
accepted October 12, 2004.
This article has been cited by other articles:
|
|
|
|
 
H.-J. Wang, A.-R. Wan, and G.-Y. Jauh
An Actin-Binding Protein, LlLIM1, Mediates Calcium and Hydrogen Regulation of Actin Dynamics in Pollen Tubes
Plant Physiology,
August 1, 2008;
147(4):
1619 - 1636.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. A. Sparkes, N. A. Teanby, and C. Hawes
Truncated myosin XI tail fusions inhibit peroxisome, Golgi, and mitochondrial movement in tobacco leaf epidermal cells: a genetic tool for the next generation
J. Exp. Bot.,
June 1, 2008;
59(9):
2499 - 2512.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. A. Konopka, S. K. Backues, and S. Y. Bednarek
Dynamics of Arabidopsis Dynamin-Related Protein 1C and a Clathrin Light Chain at the Plasma Membrane
PLANT CELL,
May 1, 2008;
20(5):
1363 - 1380.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. K. Kandasamy, B. Burgos-Rivera, E. C. McKinney, D. R. Ruzicka, and R. B. Meagher
Class-Specific Interaction of Profilin and ADF Isovariants with Actin in the Regulation of Plant Development
PLANT CELL,
October 1, 2007;
19(10):
3111 - 3126.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Thomas, C. Hoffmann, M. Dieterle, M. Van Troys, C. Ampe, and A. Steinmetz
Tobacco WLIM1 Is a Novel F-Actin Binding Protein Involved in Actin Cytoskeleton Remodeling
PLANT CELL,
September 1, 2006;
18(9):
2194 - 2206.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Higaki, N. Kutsuna, E. Okubo, T. Sano, and S. Hasezawa
Actin Microfilaments Regulate Vacuolar Structures and Dynamics: Dual Observation of Actin Microfilaments and Vacuolar Membrane in Living Tobacco BY-2 Cells
Plant Cell Physiol.,
July 1, 2006;
47(7):
839 - 852.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-U. Hwang, Y. Gu, Y.-J. Lee, and Z. Yang
Oscillatory ROP GTPase Activation Leads the Oscillatory Polarized Growth of Pollen Tubes
Mol. Biol. Cell,
November 1, 2005;
16(11):
5385 - 5399.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. L. Engel, R. Holmes-Davis, and S. McCormick
Green Sperm. Identification of Male Gamete Promoters in Arabidopsis
Plant Physiology,
August 1, 2005;
138(4):
2124 - 2133.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. O. Wasteneys and Z. Yang
The Cytoskeleton Becomes Multidisciplinary
Plant Physiology,
December 1, 2004;
136(4):
3853 - 3854.
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. O. Wasteneys and Z. Yang
New Views on the Plant Cytoskeleton
Plant Physiology,
December 1, 2004;
136(4):
3884 - 3891.
[Full Text]
[PDF]
|
|
|
|
|
