A Small Novel A-Kinase Anchoring Protein (AKAP) That Localizes Specifically Protein Kinase A-Regulatory Subunit I (PKA-RI) to the Plasma Membrane

Burgers, Pepijn P.; Ma, Yuliang; Margarucci, Luigi; Mackey, Mason; Heyden, Marcel A.G. van der; Ellisman, Mark H.; Scholten, Arjen; Taylor, Susan S.; Heck, Albert J. R.

doi:10.1074/jbc.m112.395970

Cited by 74 publications

(78 citation statements)

References 48 publications

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“…Sphingosine kinase interacting protein (SKIP) was also reported to be an R1-specific AKAP (56,57). Another small myristoylated and palmitoylated AKAP was also shown to be RI-specific and is recruited to the plasma membrane (58). R1␣ was also shown to be important for cell migration in response to ␣4 integrin, and recruitment of PKARIa also did not involve a canonical AKAP mechanism (59 -61).…”

Section: P-rex1 Dh-ph Catalytic Module Is Also Inhibited By the C-termentioning

confidence: 99%

Protein Kinase A (PKA) Type I Interacts with P-Rex1, a Rac Guanine Nucleotide Exchange Factor

Chávez-Vargas¹,

Adame-García

Cervantes‐Villagrana³

et al. 2016

Journal of Biological Chemistry

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Morphology of migrating cells is regulated by Rho GTPases and fine-tuned by protein interactions and phosphorylation.PKA affects cell migration potentially through spatiotemporal interactions with regulators of Rho GTPases. Here we show that the endogenous regulatory (R) subunit of type I PKA interacts with P-Rex1, a Rac guanine nucleotide exchange factor that integrates chemotactic signals. Type I PKA holoenzyme interacts with P-Rex1 PDZ domains via the CNB B domain of RI␣, which when expressed by itself facilitates endothelial cell migration. P-Rex1 activation localizes PKA to the cell periphery, whereas stimulation of PKA phosphorylates P-Rex1 and prevents its activation in cells responding to SDF-1 (stromal cellderived factor 1). The P-Rex1 DEP 1 domain is phosphorylated at Ser-436, which inhibits the DH-PH catalytic cassette by direct interaction. In addition, the P-Rex1 C terminus is indirectly targeted by PKA, promoting inhibitory interactions independently of the DEP 1 -PDZ 2 region. A P-Rex1 S436A mutant construct shows increased RacGEF activity and prevents the inhibitory effect of forskolin on sphingosine 1-phosphate-dependent endothelial cell migration. Altogether, these results support the idea that P-Rex1 contributes to the spatiotemporal localization of type I PKA, which tightly regulates this guanine exchange factor by a multistep mechanism, initiated by interaction with the PDZ domains of P-Rex1 followed by direct phosphorylation at the first DEP domain and putatively indirect regulation of the C terminus, thus promoting inhibitory intramolecular interactions. This reciprocal regulation between PKA and P-Rex1 might represent a key node of integration by which chemotactic signaling is fine-tuned by PKA.Rho guanine exchange factors (RhoGEFs) 4 are mechanistically linked to fundamental cellular processes, such as migration, adhesion, and morphogenesis. Based on their ability to integrate signaling inputs that result in the activation of Rho GTPases, RhoGEFs indirectly contribute to establish nucleation sites for actin polymerization, thus exerting a tight control on cytoskeleton dynamics (1, 2). RhoGEFs can also play a role as scaffolds of different signaling cascades, thus representing regulatory nodes for spatial and temporal control of signaling (3). These fundamental processes are fine-tuned by phosphorylation of RhoGEFs. However, the complex interplay between RhoGEFs and kinases is not completely understood; relevant examples include RhoGEFs known to be activated or inhibited by phosphorylation, some of them by PKA (4, 5).The ability of PKA to modulate the activity of RhoGEFs might be further facilitated by direct interactions that would also influence the subcellular localization of PKA and its accessibility to specific substrates. The paradigmatic example of this situation is AKAP-Lbc, which is a RhoGEF activated by G 12 -coupled receptors that promotes polymerization of actin filaments downstream of Rho and also interacts with multiple kinases regulating mitogenic inputs (6, 7). AKAP-Lbc ...

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Section: P-rex1 Dh-ph Catalytic Module Is Also Inhibited By the C-termentioning

confidence: 99%

Protein Kinase A (PKA) Type I Interacts with P-Rex1, a Rac Guanine Nucleotide Exchange Factor

Chávez-Vargas¹,

Adame-García

Cervantes‐Villagrana³

et al. 2016

Journal of Biological Chemistry

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“…Selectivity-Although the majority of anchoring proteins preferentially interact with type II PKA, a few type I PKA-selective AKAPs have also been reported (2,(43)(44)(45). Another subset, designated dual function AKAPs, can interact with either RI or RII (46,47).…”

Section: Pka Binding Sites On Akap220 Have Differential Isoformmentioning

confidence: 99%

Protein Kinase A Opposes the Phosphorylation-dependent Recruitment of Glycogen Synthase Kinase 3β to A-kinase Anchoring Protein 220

Whiting

Nygren

Tunquist

et al. 2015

Journal of Biological Chemistry

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Background: AKAPs integrate intracellular signals by sequestering PKA with other kinases. Results: Phosphorylation of Thr-1132 on AKAP220 initiates GSK3␤ recruitment, and PKA activity drives the release of GSK3␤ from the complex. Conclusion: Cross-talk between PKA and GSK3␤ is optimized in the context of AKAP220 multienzyme complexes. Significance: Signal responsive assembly of enzyme complexes may represent a general mechanism to diversify transduction through AKAPs.

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“…MiniSOG is not only a CALI reagent. It has also been used in correlative fluorescence and electron microscopy, in combination with diaminobenzidinewhich forms osmiophilic compounds upon reaction with singlet oxygen -and, particularly, in the field of neuroscience, has enabled the detection of barely visible objects (Boassa et al, 2013;Burgers et al, 2012;Ludwig et al, 2013;Shu, et al, 2011).…”

Section: Cali With Minisogmentioning

confidence: 99%

Chromophore-assisted laser inactivation – towards a spatiotemporal–functional analysis of proteins, and the ablation of chromatin, organelle and cell function

Sano

Watanabe

Matsunaga

2014

Journal of Cell Science

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Chromophore-assisted laser or light inactivation (CALI) has been employed as a promising technique to achieve spatiotemporal knockdown or loss-of-function of target molecules in situ. CALI is performed using photosensitizers as generators of reactive oxygen species (ROS). There are two CALI approaches that use either transgenic tags with chemical photosensitizers, or genetically encoded fluorescent protein fusions. Using spatially restricted microscopy illumination, CALI can address questions regarding, for example, protein isoforms, subcellular localization or phase-specific analyses of multifunctional proteins that other knockdown approaches, such as RNA interference or treatment with chemicals, cannot. Furthermore, rescue experiments can clarify the phenotypic capabilities of CALI after the depletion of endogenous targets. CALI can also provide information about individual events that are involved in the function of a target protein and highlight them in multifactorial events. Beyond functional analysis of proteins, CALI of nuclear proteins can be performed to induce cell cycle arrest, chromatin-or locus-specific DNA damage. Even at organelle levelsuch as in mitochondria, the plasma membrane or lysosomes -CALI can trigger cell death. Moreover, CALI has emerged as an optogenetic tool to switch off signaling pathways, including the optical depletion of individual neurons. In this Commentary, we review recent applications of CALI and discuss the utility and effective use of CALI to address open questions in cell biology.

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A Small Novel A-Kinase Anchoring Protein (AKAP) That Localizes Specifically Protein Kinase A-Regulatory Subunit I (PKA-RI) to the Plasma Membrane

Cited by 74 publications

References 48 publications

Protein Kinase A (PKA) Type I Interacts with P-Rex1, a Rac Guanine Nucleotide Exchange Factor

Protein Kinase A (PKA) Type I Interacts with P-Rex1, a Rac Guanine Nucleotide Exchange Factor

Protein Kinase A Opposes the Phosphorylation-dependent Recruitment of Glycogen Synthase Kinase 3β to A-kinase Anchoring Protein 220

Chromophore-assisted laser inactivation – towards a spatiotemporal–functional analysis of proteins, and the ablation of chromatin, organelle and cell function

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