Feedback inhibition of cAMP effector signaling by a chaperone-assisted ubiquitin system

Rinaldi, Laura; Donne, Rossella Delle; Catalanotti, Bruno; Torres‐Quesada, Omar; Enzler, Florian; Moraca, Federica; Nisticò, Robert; Chiuso, Francesco; Piccinin, Sonia; Bachmann, Verena; Lindner, Herbert; Garbi, Corrado; Scorziello, Antonella; Russo, Nicola Antonino; Synofzik, Matthis; Stelzl, Ulrich; Annunziato, Lucio; Stefan, Eduard; Feliciello, Antonio

doi:10.1038/s41467-019-10037-y

Cited by 32 publications

(28 citation statements)

References 70 publications

(75 reference statements)

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“…We had previously shown that ubiquitin-mediated degradation of PKA subunits is a way to control PKA activity [66]. In addition, STUB1 can mediate the ubiquitination and thus degradation of catalytic PKA subunits [67,68]. However, in MCD4 cells the knockdown of STUB1 did not alter PKA subunit abundance ( Figure 5C).…”

Section: A Cytosolicmentioning

confidence: 83%

Cyclin-Dependent Kinase 18 Controls Trafficking of Aquaporin-2 and Its Abundance through Ubiquitin Ligase STUB1, Which Functions as an AKAP

Dema

Faust

Lazarow

et al. 2020

Cells

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Arginine-vasopressin (AVP) facilitates water reabsorption in renal collecting duct principal cells through regulation of the water channel aquaporin-2 (AQP2). The hormone binds to vasopressin V2 receptors (V2R) on the surface of the cells and stimulates cAMP synthesis. The cAMP activates protein kinase A (PKA), which initiates signaling that causes an accumulation of AQP2 in the plasma membrane of the cells facilitating water reabsorption from primary urine and fine-tuning of body water homeostasis. AVP-mediated PKA activation also causes an increase in the AQP2 protein abundance through a mechanism that involves dephosphorylation of AQP2 at serine 261 and a decrease in its poly-ubiquitination. However, the signaling downstream of PKA that controls the localization and abundance of AQP2 is incompletely understood. We carried out an siRNA screen targeting 719 kinase-related genes, representing the majority of the kinases of the human genome and analyzed the effect of the knockdown on AQP2 by high-content imaging and biochemical approaches. The screening identified 13 hits whose knockdown inhibited the AQP2 accumulation in the plasma membrane. Amongst the candidates was the so far hardly characterized cyclin-dependent kinase 18 (CDK18). Our further analysis revealed a hitherto unrecognized signalosome comprising CDK18, an E3 ubiquitin ligase, STUB1 (CHIP), PKA and AQP2 that controls the localization and abundance of AQP2. CDK18 controls AQP2 through phosphorylation at serine 261 and STUB1-mediated ubiquitination. STUB1 functions as an A-kinase anchoring protein (AKAP) tethering PKA to the protein complex and bridging AQP2 and CDK18. The modulation of the protein complex may lead to novel concepts for the treatment of disorders which are caused or are associated with dysregulated AQP2 and for which a satisfactory treatment is not available, e.g., hyponatremia, liver cirrhosis, diabetes insipidus, ADPKD or heart failure.

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Section: A Cytosolicmentioning

confidence: 83%

Cyclin-Dependent Kinase 18 Controls Trafficking of Aquaporin-2 and Its Abundance through Ubiquitin Ligase STUB1, Which Functions as an AKAP

Dema

Faust

Lazarow

et al. 2020

Cells

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“…Blockers of PKA signaling cause the inhibition of this trafficking pathway, a process that is accompanied by the swelling of the Golgi apparatus [32,49]. It has been shown that the catalytic PKA subunit (PKAc) is ubiquitinated by the CHIP E3 ligase, resulting in proteasomal degradation of PKAc and signaling shutdown [86]. UVRAG mediates the interaction of β´-COP with ER tethers and COP-I coatomer for efficient fusion of retrograde vesicles to the ER, a crucial step during Golgi-to-ER retrograde transport [53].…”

Section: Discussionmentioning

confidence: 99%

The Proteasomal Deubiquitinating Enzyme PSMD14 Regulates Macroautophagy by Controlling Golgi-to-ER Retrograde Transport

Bustamante

Cereceda

González

et al. 2020

Cells

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Ubiquitination regulates several biological processes, however the role of specific members of the ubiquitinome on intracellular membrane trafficking is not yet fully understood. Here, we search for ubiquitin-related genes implicated in protein membrane trafficking performing a High-Content siRNA Screening including 1187 genes of the human “ubiquitinome” using amyloid precursor protein (APP) as a reporter. We identified the deubiquitinating enzyme PSMD14, a subunit of the 19S regulatory particle of the proteasome, specific for K63-Ub chains in cells, as a novel regulator of Golgi-to-endoplasmic reticulum (ER) retrograde transport. Silencing or pharmacological inhibition of PSMD14 with Capzimin (CZM) caused a robust increase in APP levels at the Golgi apparatus and the swelling of this organelle. We showed that this phenotype is the result of rapid inhibition of Golgi-to-ER retrograde transport, a pathway implicated in the early steps of the autophagosomal formation. Indeed, we observed that inhibition of PSMD14 with CZM acts as a potent blocker of macroautophagy by a mechanism related to the retention of Atg9A and Rab1A at the Golgi apparatus. As pharmacological inhibition of the proteolytic core of the 20S proteasome did not recapitulate these effects, we concluded that PSMD14, and the K63-Ub chains, act as a crucial regulatory factor for macroautophagy by controlling Golgi-to-ER retrograde transport.

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“…Blockers of PKA signaling cause inhibition of this trafficking pathway, a process that is accompanied by the swelling of the Golgi apparatus 44, 45 . It has been shown that the catalytic PKA subunit (PKAc) is ubiquitinated by the CHIP E3 ligase, resulting in proteasomal degradation of PKAc and signaling shutdown 77 . UVRAG mediates the interaction of β’-COP with ER tethers and COP-I coatomer for efficient fusion of retrograde vesicles to the ER, a crucial step during Golgi-to-ER retrograde transport 49 .…”

Section: Discussionmentioning

confidence: 99%

The proteasomal deubiquitinating enzyme PSMD14 regulates macroautophagy by controlling Golgi-to-ER retrograde transport

Bustamante

Cereceda

González

et al. 2020

Preprint

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31Ubiquitination regulates several biological processes. Here, we search for ubiquitin-related 32 genes implicated in protein membrane trafficking performing a High-Content siRNA 33 Screening including 1,187 genes of the human "ubiquitinome" using Amyloid Precursor 34 Protein (APP) as a reporter. We identified the deubiquitinating enzyme PSMD14, a subunit 35 of the 19S regulatory particle of the proteasome, specific for K63-Ub chains in cells, as a 36novel key regulator of Golgi-to-endoplasmic reticulum (ER) retrograde transport. Silencing 37 or pharmacological inhibition of PSMD14 caused a robust and rapid inhibition of Golgi-to-38 ER retrograde transport which leads to a potent blockage of macroautophagy by a 39 mechanism associated with the retention of Atg9A and Rab1A at the Golgi apparatus. 40Because pharmacological inhibition of the proteolytic core of the 20S proteasome did not 41 recapitulate these effects, we concluded that PSMD14, and their K-63-Ub chains, act as a 42 crucial regulator factor for macroautophagy by controlling Golgi-to-ER retrograde 43 transport. 44

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Feedback inhibition of cAMP effector signaling by a chaperone-assisted ubiquitin system

Cited by 32 publications

References 70 publications

Cyclin-Dependent Kinase 18 Controls Trafficking of Aquaporin-2 and Its Abundance through Ubiquitin Ligase STUB1, Which Functions as an AKAP

Cyclin-Dependent Kinase 18 Controls Trafficking of Aquaporin-2 and Its Abundance through Ubiquitin Ligase STUB1, Which Functions as an AKAP

The Proteasomal Deubiquitinating Enzyme PSMD14 Regulates Macroautophagy by Controlling Golgi-to-ER Retrograde Transport

The proteasomal deubiquitinating enzyme PSMD14 regulates macroautophagy by controlling Golgi-to-ER retrograde transport

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