Rho GTPase isoforms in cell motility: Don't fret, we have FRET.
Rho GTPase isoforms in cell motility: Don't fret, we have FRET.
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DOI：
10.4161/cam.29712
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发表时间：
2014
影响因子：
3.2
通讯作者：
Hodgson L
中科院分区：
文献类型：
--
作者：
Donnelly SK;Bravo-Cordero JJ;Hodgson L
p21 小 GTP 酶的 Rho 家族与基于肌动蛋白的运动机制的调节直接相关，并在细胞迁移的控制中发挥关键作用。除了最初和最充分表征的规范 Rho GTP 酶 RhoA、Rac1 和 Cdc42 之外，这些关键蛋白的许多亚型已被鉴定并显示在调节各种细胞运动过程中具有特定作用。解决这些异构体特异性效应的主要困难在于，异构体通常含有高度相似的一级氨基酸序列，因此能够与相同的上游调节因子和下游效应靶标相互作用。在这里，我们将介绍每个 GTPase 亚家族的主要成员，并讨论基于荧光共振能量转移的探针的设计和应用的最新进展，这些探针处于可直接探测活单细胞中这些蛋白质的差异时空激活动力学的最前沿技术。目前，可以特异性检测活细胞中 RhoA 与 RhoC 同工型以及 Cdc42 与 TC-10 同工型的激活状态。显然，仍然需要付出额外的努力来生产能够检测其他 Rho GTP 酶亚型（包括 RhoB、Rac2/3、RhoG 等）的生物传感器系统。通过这些努力，我们将揭示这些几乎相同的蛋白质在活细胞中的亚型特异性作用，这显然是 Rho GTP 酶生物学的一个尚未完全认识的重要领域。
The Rho-family of p21 small GTPases are directly linked to the regulation of actin-based motile machinery and play a key role in the control of cell migration. Aside from the original and most well-characterized canonical Rho GTPases RhoA, Rac1, and Cdc42, numerous isoforms of these key proteins have been identified and shown to have specific roles in regulating various cellular motility processes. The major difficulty in addressing these isoform-specific effects is that isoforms typically contain highly similar primary amino acid sequences and thus are able to interact with the same upstream regulators and the downstream effector targets. Here, we will introduce the major members of each GTPase subfamily and discuss recent advances in the design and application of fluorescent resonance energy transfer-based probes, which are at the forefront of the technologies available to directly probe the differential, spatiotemporal activation dynamics of these proteins in live single cells. Currently, it is possible to specifically detect the activation status of RhoA vs. RhoC isoforms, as well as Cdc42 vs. TC-10 isoforms in living cells. Clearly, additional efforts are still required to produce biosensor systems capable of detecting other isoforms of Rho GTPases including RhoB, Rac2/3, RhoG, etc. Through such efforts, we will uncover the isoform-specific roles of these near-identical proteins in living cells, clearly an important area of the Rho GTPase biology that is not yet fully appreciated.