Direct Interaction of p21-Activated Kinase 4 with PDZ-RhoGEF, a G Protein-linked Rho Guanine Exchange Factor

Barac, Ana; Basile, John R.; Vázquez‐Prado, José; Gao, Yuan; Zheng, Yi; Gutkind, J. Silvio

doi:10.1074/jbc.m309579200

Cited by 64 publications

(49 citation statements)

References 34 publications

(45 reference statements)

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“…This is also consistent with our findings that PAK4 kinase activity leads to dissolution of stress fibers through phosphorylation of GEF-H1 in NIH-3T3 cells. Our findings related to antagonism of GEF-H1-dependent Rho activity, which is dependent upon PAK4 recruitment and kinase activity, are consistent with recent studies showing that PAK4 can antagonize Rho activity through direct interaction with RhoGEF (Barac et al, 2004).…”

Section: Discussionsupporting

confidence: 79%

PAK4 mediates morphological changes through the regulation of GEF-H1

Callow¹,

Zozulya²,

Gishizky³

et al. 2005

Journal of Cell Science

146

127

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Precise spatial and temporal regulation of Rho GTPases is required in controlling F-actin-based changes in cell morphology. The molecular mechanisms through which microtubules (MTs) modulate the activity of RhoGTPases and regulate the actin cytoskeleton are unclear. Here we show that p21-activated-kinase 4 (PAK4) mediates morphological changes through its association with the Rho-family guanine nucleotide exchange factor (GEF), GEF-H1. We show that this association is dependent upon a novel GEF-H1 interaction domain (GID) within PAK4. Further, we show that PAK4-mediated phosphorylation of Ser810 acts as a switch to block GEF-H1-dependent stress fiber formation while promoting the formation of lamellipodia in NIH-3T3 cells. We found that the endogenous PAK4-GEF-H1 complex associates with MTs and that PAK4 phosphorylation of MT-bound GEF-H1 releases it into the cytoplasm of NIH-3T3 cells, which coincides with the dissolution of stress fibers. Our observations propose a novel role for PAK4 in GEF-H1-dependent crosstalk between MTs and the actin cytoskeleton.

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Section: Discussionsupporting

confidence: 79%

PAK4 mediates morphological changes through the regulation of GEF-H1

Callow¹,

Zozulya²,

Gishizky³

et al. 2005

Journal of Cell Science

146

127

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“…Instead, these two proteins have a positive functional interaction in S. pombe cells: gain of Rga8 function exacerbates phenotypes caused by gain of Rho1 function and vice versa, and both Rho1 and Rga8 antagonize Pak1 function . This linkage between a member of the Pak family and a regulator of Rho is not unique to fission yeast, because two groups have recently reported that both group A and group B mammalian Paks phosphorylate and thereby alter the localization and/or activity of additional Rho-GEFs (Barac et al, 2004;Zenke et al, 2004).…”

Section: Pak1/shk1mentioning

confidence: 99%

“…Expression of activated Pak4 in fibroblasts decreases adhesion to the extracellular matrix and promotes proliferation, leading to anchorage-independent growth and increased cell migration (Callow et al, 2002;Qu et al, 2001). Pak4 substrates include the cytoskeletal regulatory kinase LIMK1, the pro-apoptotic protein BAD and probably PDZRho-GEF (Barac et al, 2004;Dan, C. et al, 2001;Gnesutta et al, 2001).…”

Section: Group B Paksmentioning

confidence: 99%

The genetics of Pak

Hofmann

Шепелев

Chernoff

2004

Journal of Cell Science

224

234

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p21-activated kinases (Paks) are a highly conserved family of enzymes that bind to and are activated by small GTPases of the Cdc42 and Rac families. With the notable exception of plants, nearly all eukaryotes encode one or more Pak genes, indicating an ancient origin and important function for this family of enzymes. Genetic approaches in many different experimental systems, ranging from yeast to mice, are beginning to decipher the different functions of Paks. Although some of these functions are unique to a given organism, certain common themes have emerged, such as the activation of mitogen-activated protein kinase (MAPK) cascades and the regulation of cytoskeletal structure through effects on the actin and tubulin cytoskeletons.

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“…These GEFs may then interact through other regions with membrane components, such as membrane phospholipids through their PH domain, thus either causing a conformational change in the DH domain that then becomes activated, or adopting an orientation with respect to the lipid bilayer that facilitates the interaction of the DH domain with membrane-bound Rho. This membrane interaction might be also required for the regulatory properties of tyrosine or serine/threonine protein kinases that modulate RGL-RhoGEFs, such as focal adhesion kinase and PAK4 (21,37), respectively. These, as well as other possibilities are under current investigation.…”

Section: Figmentioning

confidence: 99%

Chimeric Gαi2/Gα13 Proteins Reveal the Structural Requirements for the Binding and Activation of the RGS-like (RGL)-containing Rho Guanine Nucleotide Exchange Factors (GEFs) by Gα13

Vázquez‐Prado

Miyazaki

Castellone

et al. 2004

Journal of Biological Chemistry

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The ␣-subunit of G proteins of the G 12/13 family stimulate Rho by their direct binding to the RGS-like (RGL) domain of a family of Rho guanine nucleotide exchange factors (RGL-RhoGEFs) that includes PDZ-RhoGEF (PRG), p115RhoGEF, and LARG, thereby regulating cellular functions as diverse as shape and movement, gene expression, and normal and aberrant cell growth. The structural features determining the ability of G␣ 12/13 to bind RGL domains and the mechanism by which this association results in the activation of RGL-RhoGEFs are still poorly understood. Here, we explored the structural requirements for the functional interaction between G␣ 13 and RGL-RhoGEFs based on the structure of RGL domains and their similarity with the area by which RGS4 binds the switch region of G␣ i proteins. Using G␣ i2 , which does not bind RGL domains, as the backbone in which G␣ 13 sequences were swapped or mutated, we observed that the switch region of G␣ 13 is strictly necessary to bind PRG, and specific residues were identified that are critical for this association, likely by contributing to the binding surface. Surprisingly, the switch region of G␣ 13 was not sufficient to bind RGL domains, but instead most of its GTPase domain is required. Furthermore, membrane localization of G␣ 13 and chimeric G␣ i2 proteins was also necessary for Rho activation. These findings revealed the structural features by which G␣ 13 interacts with RGL domains and suggest that molecular interactions occurring at the level of the plasma membrane are required for the functional activation of the RGL-containing family of RhoGEFs.Rho GTPases, which include Rho, Rac, and Cdc42, play a central role in the regulation of a number of basic cellular events such as cell movement and changes in cell shape, as well as in the control of gene expression regulation and cell growth (1). These GTP-binding proteins act as molecular switches that are inactive in their GDP-bound form, and upon exchange of GDP for GTP, they adopt an active conformation in which they can interact with their specific effector molecules, thereby affecting their localization and/or activity (1-3). This nucleotide exchange is promoted by a large family of guanine nucleotide exchange factors (GEFs), 1 the vast majority of which are characterized by the presence of a dbl-homology (DH) and pleckstrin homology (PH) domain (2, 4). These GEFs also exhibit a number of additional regulatory regions by which they are strictly controlled by a diverse array of upstream signaling pathways, including those initiated by cell adhesion molecules, tyrosine kinase growth factor receptors, as well as by G proteincoupled receptors (GPCRs) (2, 5).In particular for Rho, this GTPase participates in many physiological and pathological processes that involve the activation of GPCRs. For example, GPCRs such as those for thrombin and lysophosphatidic acid (LPA) promote cytoskeletal changes and expression from serum responsive element (SRE)-regulated genes by activating Rho (6, 7). Rho also participates in platelet aggre...

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Direct Interaction of p21-Activated Kinase 4 with PDZ-RhoGEF, a G Protein-linked Rho Guanine Exchange Factor

Cited by 64 publications

References 34 publications

PAK4 mediates morphological changes through the regulation of GEF-H1

PAK4 mediates morphological changes through the regulation of GEF-H1

The genetics of Pak

Chimeric Gαi2/Gα13 Proteins Reveal the Structural Requirements for the Binding and Activation of the RGS-like (RGL)-containing Rho Guanine Nucleotide Exchange Factors (GEFs) by Gα13

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