Green fluorescent protein fusions to Arabidopsis fimbrin 1 for spatio-temporal imaging of F-actin dynamics in roots
Green fluorescent protein fusions to Arabidopsis fimbrin 1 for spatio-temporal imaging of F-actin dynamics in roots
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DOI：
10.1002/cm.20024
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发表时间：
2004-10-01
影响因子：
--
通讯作者：
Blancaflor, EB
中科院分区：
文献类型：
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作者：
Wang, YS;Motes, CM;Blancaflor, EB
绿色荧光蛋白（GFP）与微管或肌动蛋白丝（F-actin）结合蛋白融合的可视化为植物发育过程中细胞骨架的功能提供了新的见解。对于肌动蛋白的研究，GFP融合talin已被最常用的报告。虽然GFP-塔林已允许在体内F-肌动蛋白成像在各种植物细胞中，其效用在监测F-肌动蛋白在稳定转化的植物是有限的，特别是在发展中的根，其中有趣的肌动蛋白依赖的细胞过程正在发生。在这项研究中，我们创建了多种GFP融合拟南芥Fimm 1（AtFim 1），以探索其在根细胞中的体内F-肌动蛋白成像的效用，并更好地了解AtFim 1在活植物细胞中的肌动蛋白结合特性。翻译融合的GFP全长AtFim 1或一些截断的AtFim 1变体显示丝状标记在瞬时表达试验。一种截短的fimbrin-GFP融合体能够标记稳定转化的拟南芥根中的不同细丝。由这种结构修饰的花丝在生长根毛和伸长根细胞中具有高度动态性，并且对肌动蛋白破坏药物敏感。因此，我们在这项研究中描述的fimbrin-GFP报告提供了额外的工具，研究肌动蛋白细胞骨架在根细胞发育。此外，AtFim 1-GFP的本地化提供了深入了解这类肌动蛋白交联蛋白在发展中的根的肌动蛋白组织的调节。(C)2004 Wiley-Liss，Inc.
The visualization of green fluorescent protein (GFP) fusions with microtubule or actin filament (F-actin) binding proteins has provided new insights into the function of the cytoskeleton during plant development. For studies on actin, GFP fusions to talin have been the most generally used reporters. Although GFP-Talin has allowed in vivo F-actin imaging in a variety of plant cells, its utility in monitoring F-actin in stably transformed plants is limited particularly in developing roots where interesting actin dependent cell processes are occurring. In this study, we created a variety of GFP fusions to Arabidopsis Fimbrin 1 (AtFim1) to explore their utility for in vivo F-actin imaging in root cells and to better understand the actin binding properties of AtFim1 in living plant cells. Translational fusions of GFP to full-length AtFim1 or to some truncated variants of AtFim1 showed filamentous labeling in transient expression assays. One truncated fimbrin-GFP fusion was capable of labeling distinct filaments in stably transformed Arabidopsis roots. The filaments decorated by this construct were highly dynamic in growing root hairs and elongating root cells and were sensitive to actin disrupring drugs. Therefore, the fimbrin-GFP reporters we describe in this study provide additional tools for studying the actin cytoskeleton during root cell development. Moreover, the localization of AtFim1-GFP offers insights into the regulation of actin organization in developing roots by this class of actin cross-linking proteins. (C) 2004 Wiley-Liss, Inc.