Rho GTPase对细胞骨架及信号通路的多重影响

Rho GTPase最基本的功能是结合和水解鸟嘌呤核苷酸，目前已从晡乳动物中分离出16种不同的Rho GTPases，其中以Rho、Rac和Cdc42最为人们关注。研究发现Rho GTPases参与基因转录、细胞周期进程的调控及多条信号通路的调节，与细胞凋亡、肿瘤侵润及细胞骨架构成关系密切。现普遍认为Rho GTPases是调节细胞功能的一类重要蛋白分子，越来越多的Rho家族成员及其调控的蛋白数量逐渐被发现和认识。     

Rho GTPases与肿瘤

Rho GTPases参与调控细胞的多种关键生物学行为,特别是细胞的生长、细胞骨架的形成、转录调节等生物学过程. 在肿瘤的发生发展中Rho GTPases也扮演了重要的角色．本文将回顾Rho GTPases的调控（包括经典及非经典调控方式）及其关键成员（RhoA、Cdc42及Rac1）与临床肿瘤的研究进展,特别是它们参与调控肿瘤的增殖、迁移、侵袭、凋亡等恶性生物学行为,从而为研发靶向Rho GTPases的小分子/基因药物了奠定基础．     

Rho GTPases及其在口腔领域的新应用

Rho GTPases是重要的信号转导分子,参与多种重要的细胞生命活动,如肌动蛋白细胞骨架的重构、细胞黏附、细胞运动、囊泡运输、基因表达和细胞周期的调控等.调节这些生物信号的转导通路非常复杂,因此,Rho GTPases早已成为研究的热点.最新的研究进展集中在描述Rho GTPases具体在细胞的哪个部位发生反应与参与通路的具体的分子及新功能等,同时细胞分子实验已经证实Rho GTPases在肌动蛋白细胞骨架的组装、细胞粘附、细胞运动、和基因表达等方面的作用与临床上多种口腔疾病,如正畸,牙周疾病的发生有密切关系.因此,对Rho GTPases的研究可能为口腔领域正畸,牙周病的治疗提供新的思路.     

Rho GTPases与骨髓间充质干细胞的迁移和分化

Rho家族小分子鸟苷三磷酸酶（small GTPases of Rho family,Rho GTPases）是调节细胞许多生理病理活动的关键分子开关,参与细胞骨架、基因转录、细胞周期进程、细胞黏附的调控及多条信号通路的调节。骨髓间充质干细胞（bone marrow-derived mesenchymal stem cells,MSCs）是一类具有自我更新和多向分化潜能的特殊细胞,通过增殖、迁移、分化等途径参与机体损伤组织的修复过程。研究表明,Rho GT-Pases在MSCs迁移、分化等过程中起着重要的调节作用。     

Rho小G蛋白相关激酶的结构与功能

Rho小G蛋白家族是Ras超家族成员之一,人类Rho小G蛋白包括20个成员,研究最清楚的有RhoA、Rac1和Cdc42。Rho小G蛋白参与了诸如细胞骨架调节、细胞移动、细胞增殖、细胞周期调控等重要的生物学过程。在这些生物学过程的调节中,Rho小G蛋白的下游效应蛋白质如蛋白激酶（p21-activated kinase,PAK）、ROCK（Rho-kinase）、PKN（protein kinase novel）和MRCK（myotonin-related Cdc42-binding kinase）发挥了不可或缺的作用。迄今研究发现,PAK可调节细胞骨架动力学和细胞运动,另外,PAK通过MAPK（mitogen-activated protein kinases）参与转录、细胞凋亡和幸存通路及细胞周期进程;ROCK与肌动蛋白应力纤维介导黏附复合物的形成及与细胞周期进程的调节有关;哺乳动物的PKN与RhoA/B/C相互作用介导细胞骨架调节;MRCK与细胞骨架重排、细胞核转动、微管组织中心再定位、细胞移动和癌细胞侵袭等有关。该文简要介绍Rho小G蛋白下游激酶PAK、ROCK、PKN和MRCK的结构及其在细胞骨架调节中的功能,重点总结它们在真核细胞周期调控中的作用,尤其是在癌细胞周期进程中所发挥的作用,为寻找癌症治疗的新靶点提供理论依据。     

Rho蛋白和细胞骨架的关系

Rho蛋白作为细胞信号转导的分子开关之一,在细胞骨架动态变化中发挥着极其重要的作用。Rho蛋白对细胞骨架动态变化的调节是一个复杂的信号传递过程,涉及到Rho蛋白介导的信号通路中不同效应物间和Rho蛋白介导的多条信号通路间的相互作用。在Rho蛋白介导的信号通路中,上游调控因子、Rho蛋白、效应物在细胞中的正确定位对信号传递有着决定性的作用。     

Rho GTP酶参与肿瘤调控的下游效应蛋白

小G蛋白Rho家族是一类能结合GTP的蛋白质,在哺乳动物细胞的信号转导系统中发挥着“分子开关”样的重要作用。Rho蛋白通过其下游效应蛋白介导发挥多种生物学效应,包括调控细胞骨架重组、黏附和运动等。近年来,Rho蛋白在肿瘤细胞的增殖、分化、凋亡和转移的调控中的作用渐受重视。该文就介导RhoGTP酶调控肿瘤发生发展的几个下游蛋白作一介绍。     

睾丸间质细胞——研究自体吞噬的一种正常细胞模型

细胞在生理状态下自体吞噬出现的频率很低,很难用正常细胞来研究自体吞噬活动,一般都通过诱导自体吞噬来获得有关自体吞噬活动的资料。本实验观察了肝、肾、睾丸等组织的32种细胞,发现睾丸间质细胞中自体吞噬出现频率远远高于其他细胞,平均每100个细胞切面中可以看到25个自噬小体,从而为研究自体吞噬的过程和机理提供了一个正常细胞模型。本实验还观察到睾丸间质细胞的自体吞噬活动可分为前自噬小体、早期自噬小体和晚期自噬小体三个阶段,是一个连续的过程。前自噬小体和早期自噬小体不含溶酶体酶,只有在自噬小体与溶酶体接触后,才从后者获取溶酶体酶并将其内容物消化分解,成为晚期自噬小体。由自体吞噬所产生的残余体并不在睾丸间质细胞内积聚,而是通过胞吐作用排出细胞外。     

小G蛋白ROP的研究进展

小G蛋白(small GTPases)是近年来研究细胞信号转导过程的热点问题,包括Ras、Rab、Rho、Arf和Ran5个亚家族,其中ROP蛋白是Rho家族成员,为植物特有,在调控细胞生长、发育及调节植物对环境响应等各方面起重要作用.对ROP蛋白的活性调节和功能进行了重点介绍.     

凋亡细胞清除的分子和细胞生物学机制

凋亡细胞的正确清除是细胞程序性死亡的必要环节,对生物体的器官发育和组织稳态维持起至关重要的作用.凋亡细胞的清除涉及凋亡细胞与吞噬细胞之间的相互识别、吞噬小体形成、吞噬小体与溶酶体融合、凋亡细胞降解等多个环节.基于模式动物秀丽词线虫的研究使人们对这一过程的调控机制有了系统的认识,本文将简要综述该方面的研究进展.     

Roles of Rho-family GTPases in cell polarisation and directional migration

Polarised cell migration is a tightly regulated process that occurs in tissue development, chemotaxis and wound healing. Rho-family GTPases, including Cdc42, Rac1 and RhoA, play a central role in establishing cell polarisation, which requires asymmetric and ordered distribution of the signalling molecules and the cytoskeleton. Recent advances reveal that Rho GTPases, together with phosphatidylinositol 3-kinase, contribute to asymmetric phosphatidylinositol 3,4,5-trisphosphate distribution via a positive-feedback loop. Phosphatidylinositol 3,4,5-trisphosphate thereby activates the signalling cascades to the cytoskeleton as a second messenger. Rho GTPases also capture and stabilise microtubules through their effectors (e.g. IQGAP1, mDia and Par6) near the cell cortex, leading to polarised cell morphology and directional cell migration. Thus, elucidation of the signal transduction cascades from receptors to Rho GTPases and, subsequently, from Rho GTPases to microtubules has begun.     

Rho GTPases in neuronal morphogenesis

The Rho family of small GTPases act as intracellular molecular switches that transduce signals from extracellular stimuli to the actin cytoskeleton and the nucleus. Recent evidence implicates Rho GTPases in the regulation of neuronal morphogenesis, including migration, polarity, axon growth and guidance, dendrite elaboration and plasticity, and synapse formation. Signalling pathways from membrane receptors to Rho GTPases and from Rho GTPases to the actin cytoskeleton are beginning to be discovered. Mutations in these signalling pathways have been reported in human neurological diseases, which underscores their importance in the development and function of the nervous system.     

The role of Rho GTPases and associated kinases in regulating neurite outgrowth

Neurones are highly specialised cells that can extend over great distances, enabling the complex networking of the nervous system. We are beginning to understand in detail the molecular mechanisms that control the shape of neurones during development. One family of proteins that are clearly essential are the Rho GTPases which have a pivotal role in regulating the actin cytoskeleton in all cell types. The Rho GTPases are responsible for the activation and downregulation of many downstream kinases. This review discusses individual kinases that are regulated by three members of the Rho GTPases, Rac, Rho and Cdc42 and their function during neurite outgrowth and remodelling.     

Rho GTPases in growth cone guidance

It is now well established that the small GTPases of the Rho family--Rac, Cdc42 and Rho--regulate growth cone morphology. Less clear is their role in guiding the growth cone. Do they act permissively, providing the dynamic actin structures needed for guidance? Or do they act instructively, transducing specific guidance signals? Recent studies have provided the first strong evidence for an instructive role: extracellular guidance cues can modulate Rho GTPase activities in vitro, and Rho GTPase activators function in growth cone guidance in vivo. The pathways linking Rho GTPases and the actin cytoskeleton are also rapidly coming into view, revealing further points of regulation by extracellular guidance cues. The growth cone is therefore guided by signals transduced both via and independently of Rho GTPases.     

Rho GTPases in animal cell mitosis

The Rho GTPases have been thought to influence cell morphogenesis through remodeling of the actin cytoskeleton. Consistently, downstream targets such as the mDia family of formins and the WASP family proteins induce actin nucleation and polymerization, and another set of downstream effectors, the ROCK family protein kinases, are involved in regulation of actomyosin contractility. However, evidence has now accumulated that Rho GTPases also regulate local dynamics of microtubules. The mDia family proteins, for example, function downstream of Rho to stabilize and align microtubules in interphase cells. Concomitantly, the role of Rho GTPases in animal cell division, once thought to be limited to cytokinesis, has now been shown to extend to mitosis. Recent work indicates that they may function during both spindle orientation and chromosome congression. However, their involvement is cell-type-specific, raising arguments for and against a mitotic role for Rho GTPases.     

Inactivation of small Rho GTPases by the multifunctional RTX toxin from Vibrio cholerae

Many bacterial toxins target small Rho GTPases in order to manipulate the actin cytoskeleton. The depolymerization of the actin cytoskeleton by the Vibrio cholerae RTX toxin was previously identified to be due to the unique mechanism of covalent actin cross-linking. However, identification and subsequent deletion of the actin cross-linking domain within the RTX toxin revealed that this toxin has an additional cell rounding activity. In this study, we identified that the multifunctional RTX toxin also disrupts the actin cytoskeleton by causing the inactivation of small Rho GTPases, Rho, Rac and Cdc42. Inactivation of Rho by RTX was reversible in the presence of cycloheximide and by treatment of cells with CNF1 to constitutively activate Rho. These data suggest that RTX targets Rho GTPase regulation rather than affecting Rho GTPase directly. A novel 548-amino-acid region of RTX was identified to be responsible for the toxin-induced inactivation of the Rho GTPases. This domain did not carry GAP or phosphatase activities. Overall, these data show that the RTX toxin reversibly inactivates Rho GTPases by a mechanism distinct from other Rho-modifying bacterial toxins.     

Cell migration: Rho GTPases lead the way

Rho GTPases control signal transduction pathways that link cell surface receptors to a variety of intracellular responses. They are best known as regulators of the actin cytoskeleton, but in addition they control cell polarity, gene expression, microtubule dynamics and vesicular trafficking. Through these diverse functions, Rho GTPases influence many aspects of cell behavior. This review will focus specifically on their role in cell migration.     

Integrin-mediated Signals Regulated by Members of the Rho Family of GTPases

The organization of the actin cytoskeleton can be regulated by soluble factors that trigger signal transduction events involving the Rho family of GTPases. Since adhesive interactions are also capable of organizing the actin-based cytoskeleton, we examined the role of Cdc42-, Rac-, and Rho-dependent signaling pathways in regulating the cytoskeleton during integrin-mediated adhesion and cell spreading using dominant-inhibitory mutants of these GTPases. When Rat1 cells initially adhere to the extracellular matrix protein fibronectin, punctate focal complexes form at the cell periphery. Concomitant with focal complex formation, we observed some phosphorylation of the focal adhesion kinase (FAK) and Src, which occurred independently of Rho family GTPases. However, subsequent phosphorylation of FAK and paxillin occurs in a Rho-dependent manner. Moreover, we found Rho dependence of the assembly of large focal adhesions from which actin stress fibers radiate. Initial adhesion to fibronectin also stimulates membrane ruffling; we show that this ruffling is independent of Rho but is dependent on both Cdc42 and Rac. Furthermore, we observed that Cdc42 controls the integrin-dependent activation of extracellular signal–regulated kinase 2 and of Akt, a kinase whose activity has been demonstrated to be dependent on phosphatidylinositol (PI) 3-kinase. Since Rac-dependent membrane ruffling can be stimulated by PI 3-kinase, it appears that Cdc42, PI 3-kinase, and Rac lie on a distinct pathway that regulates adhesion-induced membrane ruffling. In contrast to the differential regulation of integrin-mediated signaling by Cdc42, Rac, and Rho, we observed that all three GTPases regulate cell spreading, an event that may indirectly control cellular architecture. Therefore, several separable signaling pathways regulated by different members of the Rho family of GTPases converge to control adhesion-dependent changes in the organization of the cytoskeleton, changes that regulate cell morphology and behavior.