Structural Basis for the 14-3-3 Protein-dependent Inhibition of the Regulator of G Protein Signaling 3 (RGS3) Function

Rezabkova, Lenka; Man, Petr; Novák, Petr; Heřman, Petr; Večeř, Jaroslav; Obšilová, Veronika; Obšil, Tomáš

doi:10.1074/jbc.m111.273573

Cited by 25 publications

(30 citation statements)

References 56 publications

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“…This could be attributed to a bound fraction of DANS-TRE sensing lower solvation and decreased microenvironmental polarity in the binding pocket of the complex. The value is consistent with values commonly found for the dansyl moiety covalently linked to globular proteins (34,35). The fractional intensity of the 14.3-ns component in the Bmh1:pNth1 1-751 (E674A) complex (4.2%) is significantly higher than in the presence of pNth1 1-751 (E674A) (0.5%) or Bmh1 alone (1.3%), where the presence of the long component likely indicates some nonspecific interaction of DANS-TRE with Bmh1.…”

Section: Resultssupporting

confidence: 75%

Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

Alblova

Smidova

Dočekal

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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The 14-3-3 proteins, a family of highly conserved scaffolding proteins ubiquitously expressed in all eukaryotic cells, interact with and regulate the function of several hundreds of partner proteins. Yeast neutral trehalases (Nth), enzymes responsible for the hydrolysis of trehalose to glucose, compared with trehalases from other organisms, possess distinct structure and regulation involving phosphorylation at multiple sites followed by binding to the 14-3-3 protein. Here we report the crystal structures of yeast Nth1 and its complex with Bmh1 (yeast 14-3-3 isoform), which, together with mutational and fluorescence studies, indicate that the binding of Nth1 by 14-3-3 triggers Nth1’s activity by enabling the proper 3D configuration of Nth1’s catalytic and calcium-binding domains relative to each other, thus stabilizing the flexible part of the active site required for catalysis. The presented structure of the Bmh1:Nth1 complex highlights the ability of 14-3-3 to modulate the structure of a multidomain binding partner and to function as an allosteric effector. Furthermore, comparison of the Bmh1:Nth1 complex structure with those of 14-3-3:serotonin N-acetyltransferase and 14-3-3:heat shock protein beta-6 complexes revealed similarities in the 3D structures of bound partner proteins, suggesting the highly conserved nature of 14-3-3 affects the structures of many client proteins.

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Section: Resultssupporting

confidence: 75%

Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

Alblova

Smidova

Dočekal

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…[37][38][39][40] 14-3-3 might interfere with the binding of RGS proteins to their G␣ targets, and recent data on RGS3 suggest that 14-3-3 binding induces a conformational change in RGS3 that is likely to affect the interaction of RGS3 and G␣. 42,43 Our studies on the interaction between RGS18 and Gi2 and Gq by co-immunoprecipitation from transfected cells showed a trend toward increased binding of the 14-3-3-deficient RGS18 mutant to Gi2 and Gq compared with wt-RGS18; however, this effect was not statistically significant (data not shown). Live microscopy of fluorescently labeled wt-RGS18 and 14-3-3-deficient mutants in transfected cells did not reveal any major difference in subcellular distribution (data not shown).…”

Section: Discussionmentioning

confidence: 61%

Regulator of G-protein signaling 18 integrates activating and inhibitory signaling in platelets

Gegenbauer

Elia

Blanco

et al. 2012

Blood

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Regulator of G-protein signaling 18 (RGS18) is a GTPase-activating protein for the G-␣-q and G-␣-i subunits of heterotrimeric Gproteins that turns off signaling by G-protein coupled receptors. RGS18 is highly expressed in platelets. In the present study, we show that the 14-3-3␥ protein binds to phosphorylated serines 49 and 218 of RGS18. Platelet activation by thrombin, thromboxane A2, or ADP stimulates the association of 14-3-3 and RGS18, probably by increasing the phosphorylation of serine 49. In contrast, treatment of platelets with prostacyclin and nitric oxide, which trigger inhibitory cyclic nucleotide signaling involving cyclic AMP-dependent protein kinase A (PKA) and cyclic GMP-dependent protein kinase I (PKGI), induces the phosphorylation of serine 216 of RGS18 and the detachment of 14-3-3. Serine 216 phosphorylation is able to block 14-3-3 binding to RGS18 even in the presence of thrombin, thromboxane A2, or ADP. 14-3-3-deficient RGS18 is more active compared with 14-3-3-bound RGS18, leading to a more pronounced inhibition of thrombin-induced release of calcium ions from intracellular stores. Therefore, PKA-and PKGI-mediated detachment of 14-3-3 activates RGS18 to block Gq-dependent calcium signaling. These findings indicate cross-talk between platelet activation and inhibition pathways at the level of RGS18 and Gq. IntroductionIn healthy vasculature, endothelial cells lining the blood vessels constantly produce and release prostacyclin (PGI 2 ) and nitric oxide (NO) into the vessel lumen. The interaction of endothelial factors with platelets plays a fundamental role in controlling hemostasis and in maintaining platelets in a resting state. Platelet inhibition by both PGI 2 and NO has been well established. The signaling pathways of both molecules result in an elevation of cyclic nucleotides that activate cyclic AMP-dependent protein kinase A (PKA) and cyclic GMP-dependent protein kinase I (PKGI). These in turn phosphorylate an unknown number of substrate proteins, resulting in reduced release of calcium ions (Ca 2ϩ ) from intracellular stores and reduced activation of G-proteins such as Rap1, ultimately leading to a block of platelet adhesion, granule release, and aggregation. PKA and PKGI have overlapping substrate specificities, which may explain the synergistic role of the 2 pathways. Few substrates have been established in platelets, among them Rap1GAP2, a GTPase-activating protein (GAP) of the small G-protein Rap1, as we have shown previously. 1 Other substrates include vasodilator-stimulated phospho-protein (VASP), heatshock protein 27 (HSP27), and LIM and SH3 domain protein (LASP), all of which regulate actin dynamics. 2,3 The IP 3 -receptor and the IP 3 -receptor-associated G-kinase substrate (IRAG) are the only PKA and/or PKGI substrates that have been shown to mediate cAMP/cGMP effects on intracellular Ca 2ϩ release. [2][3][4] Limited data are available on the specific substrates and signaling events that translate PKA/G substrate phosphorylation into platelet inhibition.Conversely, bindin...

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“…Other peptides of the Hsp70⅐ATP protein were not differentially deuterated in the presence of Tomm34, including peptide 611-638 covering the C terminus. The absence of protection of the extreme Hsp70 C terminus might be caused by the high flexibility of this region (24,62) or by side chain-mediated interactions that cannot be trapped by H/D exchange. These data suggest that the identified Hsp70 segment spanning amino acids 519 -529/533-543 represent the additional Hsp70/Tomm34 interaction interface formed exclusively in the ATP-bound conformation of Hsp70, or this region reflects allosteric changes induced by Tomm34 binding to different Hsp70⅐ATP sites.…”

Section: Hsp70 Allostery and Chaperone/co-chaperone Interactionsmentioning

confidence: 97%

Novel Entropically Driven Conformation-specific Interactions with Tomm34 Protein Modulate Hsp70 Protein Folding and ATPase Activities

Ďurech

Trčka

Man

et al. 2016

Molecular & Cellular Proteomics

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Co-chaperones containing tetratricopeptide repeat (TPR) domains enable cooperation between Hsp70 and Hsp90 to maintain cellular proteostasis. Although the details of the molecular interactions between some TPR domains and heat shock proteins are known, we describe a novel mechanism by which Tomm34 interacts with and coordinates Hsp70 activities. In contrast to the previously defined Hsp70/Hsp90-organizing protein (Hop), Tomm34 interaction is dependent on the Hsp70 chaperone cycle. Tomm34 binds Hsp70 in a complex process; anchorage of the Hsp70 C terminus by the TPR1 domain is accompanied by additional contacts formed exclusively in the ATP-bound state of Hsp70 resulting in a high affinity entropically driven interaction. Tomm34 induces structural changes in determinants within the Hsp70-lid subdomain and modulates Hsp70/Hsp40-mediated refolding and Hsp40-stimulated Hsp70 ATPase activity. Because Tomm34 recruits Hsp90 through its TPR2 domain, we propose a model in which Tomm34 enables Hsp70/Hsp90 scaffolding and influences the Hsp70 chaperone cycle, providing an additional role for co-chaperones that contain multiple TPR domains in regulating protein homeostasis. Molecular & Cellular

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Structural Basis for the 14-3-3 Protein-dependent Inhibition of the Regulator of G Protein Signaling 3 (RGS3) Function

Cited by 25 publications

References 56 publications

Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

Regulator of G-protein signaling 18 integrates activating and inhibitory signaling in platelets

Novel Entropically Driven Conformation-specific Interactions with Tomm34 Protein Modulate Hsp70 Protein Folding and ATPase Activities

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