GAPs for rho-related GTPases

doi:10.1016/0168-9525(94)90114-7

Trends in Genetics

Volume 10, Issue 12, December 1994, Pages 436-440

https://doi.org/10.1016/0168-9525(94)90114-7 Get rights and content

Abstract

Ras-related GTP-binding proteins (GTPases) of the rbo subfamily play important roles in regulating the organization of the actin cytoskeleton. A large number of multifunctional proteins that can stimulate their intrinsic GTPase activity have been identified. Here, we discuss the nature of such GTPase-activating proteins (GAPs) and their potential importance for cell signalling.

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Cited by (212)

Arabidopsis pavement cell shape formation involves spatially confined ROPGAP regulators
2022, Current Biology
In many plant species, pavement cell development relies on the coordinated formation of interdigitating lobes and indentations. Polarity signaling via the activity of antagonistic Rho-related GTPases from plants (ROPs) was implicated in pavement cell development, but the spatiotemporal regulation remained unclear. Here, we report on the role of the PLECKSTRIN HOMOLOGY GTPase ACTIVATING PROTEINS (PHGAPS) during multipolar growth in pavement cell shape establishment. Loss of function in phgap1phgap2 double mutants severely affected the shape of Arabidopsis leaf epidermal pavement cells. Predominantly, PHGAPs interacted with ROP2 and displayed a distinct and microtubule-dependent enrichment along the anticlinal cell face and transfacial boundary of pavement cell indentation regions. This localization was established upon undulation initiation and was maintained throughout the expansion of the cell. Our data suggest that PHGAP1/REN2 and PHGAP2/REN3 are key players in the establishment of ROP2 activity gradients and underscore the importance of locally controlled ROP activity for the orchestrated establishment of multipolarity in epidermal cells.
Rac-dependent feedforward autoactivation of NOX2 leads to oxidative burst
2021, Journal of Biological Chemistry
NADPH oxidase 2 (NOX2) produces the superoxide anion radical (O₂⁻), which has functions in both cell signaling and immune defense. NOX2 is a multimeric-protein complex consisting of several protein subunits including the GTPase Rac. NOX2 uniquely facilitates an oxidative burst, which is described by initially slow O₂⁻ production, which increases over time. The NOX2 oxidative burst is considered critical to immune defense because it enables expedited O₂⁻ production in response to infections. However, the mechanism of the initiation and progression of this oxidative burst and its implications for regulation of NOX2 have not been clarified. In this study, we show that the NOX2 oxidative burst is a result of autoactivation of NOX2 coupled with the redox function of Rac. NOX2 autoactivation begins when active Rac triggers NOX2 activation and the subsequent production of O₂⁻, which in turn activates redox-sensitive Rac. This activated Rac further activates NOX2, amplifying the feedforward cycle and resulting in a NOX2-mediated oxidative burst. Using mutagenesis-based kinetic and cell analyses, we show that enzymatic activation of Rac is exclusively responsible for production of the active Rac trigger that initiates NOX2 autoactivation, whereas redox-mediated Rac activation is the main driving force of NOX2 autoactivation and contributes to generation of ∼98% of the active NOX2 in cells. The results of this study provide insight into the regulation of NOX2 function, which could be used to develop therapeutics to control immune responses associated with dysregulated NOX2 oxidative bursts.
Deciphering the molecular and functional basis of RHOGAP family proteins: A systematic approach toward selective inactivation of RHO family proteins
2016, Journal of Biological Chemistry
RHO GTPase-activating proteins (RHOGAPs) are one of the major classes of regulators of the RHO-related protein family that are crucial in many cellular processes, motility, contractility, growth, differentiation, and development. Using database searches, we extracted 66 distinct human RHOGAPs, from which 57 have a common catalytic domain capable of terminating RHO protein signaling by stimulating the slow intrinsic GTP hydrolysis (GTPase) reaction. The specificity of the majority of the members of RHOGAP family is largely uncharacterized. Here, we comprehensively investigated the sequence-structure-function relationship between RHOGAPs and RHO proteins by combining our in vitro data with in silico data. The activity of 14 representatives of the RHOGAP family toward 12 RHO family proteins was determined in real time. We identified and structurally verified hot spots in the interface between RHOGAPs and RHO proteins as critical determinants for binding and catalysis. We have found that the RHOGAP domain itself is nonselective and in some cases rather inefficient under cell-free conditions. Thus, we propose that other domains of RHOGAPs confer substrate specificity and fine-tune their catalytic efficiency in cells.
Thiopurine prodrugs mediate immunosuppressive effects by interfering with Rac1 protein function
2016, Journal of Biological Chemistry
6-Thiopurine (6-TP) prodrugs include 6-thioguanine and azathioprine. Both are widely used to treat autoimmune disorders and certain cancers. This study showed that a 6-thioguanosine triphosphate (6-TGTP), converted in T-cells from 6-TP, targets Rac1 to form a disulfide adduct between 6-TGTP and the redox-sensitive GXXXXGK(S/T)C motif of Rac1. This study also showed that, despite the conservation of the catalytic activity of RhoGAP (Rho-specific GAP) on the 6-TGTP-Rac1 adduct to produce the biologically inactive 6-thioguanosine diphosphate (6-TGDP)-Rac1 adduct, RhoGEF (Rho-specific GEF) cannot exchange the 6-TGDP adducted on Rac1 with free guanine nucleotide. The biologically inactive 6-TGDP-Rac1 adduct accumulates in cells because of the ongoing combined actions of RhoGEF and RhoGAP. Because other Rho GTPases, such as RhoA and Cdc42, also possess the GXXXXGK(S/T)C motif, the proposed mechanism for the inactivation of Rac1 also applies to RhoA and Cdc42. However, previous studies have shown that CD3/CD28-stimulated T-cells contain more activated Rac1 than other Rho GTPases such as RhoA and Cdc42. Accordingly, Rac1 is the main target of 6-TP in activated T-cells. This explains the T-cell-specific Rac1-targeting therapeutic action of 6-TP that suppresses the immune response. This proposed mechanism for the action of 6-TP on Rac1 performs a critical role in demonstrating the capability to design a Rac1-targeting chemotherapeutic agent(s) for autoimmune disorders. Nevertheless, the results also suggest that the targeting action of other Rho GTPases in other organ cells, such as RhoA in vascular cells, may be linked to cytotoxicities because RhoA plays a key role in vasculature functions.
Cell type-Specific signaling function of rhoa gtpase: Lessons from mouse genetargeting
2013, Journal of Biological Chemistry
Citation Excerpt :
RhoA activity in a given cell may result from a combined effect of the various regulators expressed in the cell. In addition, whereas some GEFs and GAPs may signal specifically through RhoA, others may regulate multiple Rho GTPase family members (53, 65, 66). Thus, depending on the expression pattern of various regulators and potential cross-talk with other Rho GTPases, deletion of a given RhoA regulator may or may not result in a similar phenotype as the rhoA knock-out.
RhoA GTPase is a key intracellular regulator of actomyosin dynamics and other cell functions, including adhesion, proliferation, survival, and gene expression. Most of our knowledge of RhoA signaling function is from studies in immortalized cell lines utilizing inhibitors or dominant mutant overexpression, both of which are limited in terms of specificity, dosage, and clonal variation. Recent mouse gene targeting studies of rhoA and its regulators/effectors have revealed cell type-specific signaling mechanisms in the context of mammalian physiology. The new knowledge may present therapeutic opportunities for the rational targeting of RhoA signaling-mediated pathophysiologies.
ArhGAP15, a rac-specific gtpase-activating protein, plays a dual role in inhibiting small GTpase signaling
2013, Journal of Biological Chemistry
Signaling from small GTPases is a tightly regulated process. In this work we used a protein microarray screen to identify the Rac-specific GAP, ArhGAP15, as a substrate of the Rac effectors Pak1 and Pak2. In addition to serving as a substrate of Pak1/2, we found that ArhGAP15, via its PH domain, bound to these kinases. The association of ArhGAP15 to Pak1/2 resulted in mutual inhibition of GAP and kinase catalytic activity, respectively. Knock-down of ArhGAP15 resulted in activation of Pak1/2, both indirectly, as a result of Rac activation, and directly, as a result of disruption of the ArhGAP15/Pak complex. Our data suggest that ArhGAP15 plays a dual negative role in regulating small GTPase signaling, by acting at the level of the GTPase itself, as well interacting with its effector, Pak kinase.
Background: p21-activated kinases are effectors of Rac1. How Paks are inactivated is not well understood.
Results: We found that the Rac GAP ArhGAP15 binds to and inhibits Pak2. Conversely, Pak2 binds to, phosphorylates, and inhibits ArhGAP15.
Conclusion: ArhGAP15 and Pak2 are mutual inhibitors
Significance: These results suggest that ArhGAP15 acts at two levels: through Rac GTP hydrolysis and direct interaction with Pak.

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Trends in Genetics

ReviewGAPs for rho-related GTPases

Abstract

Curr. Opin. Genet. Dev.

Cell

Cell

Curr. Opin. Cell Biol.

J. Biol. Chem.

J. Biol. Chem.

Cell

Blood

J. Biol. Chem.

J. Biol. Chem.

J. Mol. Biol.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Cell

Cell

J. Biol. Chem.

J. Biol. Chem.

Cell

J. Biol. Chem.

Cell

Review
GAPs for rho-related GTPases