Activation of the cAMP/PKA pathway induces UT-A1 urea transporter monoubiquitination and targets it for lysosomal degradation

Su, Hua; Chen, Minguang; Sands, Jeff M.; Chen, Guangping

doi:10.1152/ajprenal.00393.2013

Cited by 14 publications

(13 citation statements)

References 37 publications

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“…On the other hand, mono‐ or short ubiquitination are involved in the endocytosis of membrane proteins, followed by sorting to the lysosome (Su et al . ; Goo et al . ).…”

Section: N‐glycosylation Mutants Proceed Into the Lysosomal Degradatimentioning

confidence: 99%

“…The proteasome pathway is mediated by protein polyubiquitination. On the other hand, mono-or short ubiquitination are involved in the endocytosis of membrane proteins, followed by sorting to the lysosome (Su et al 2013;Goo et al 2015). The agonist-induced receptor internalization of GluA1, in part, mediates lysosomal degradation (Schwarz et al 2010).…”

Section: N-glycosylation Mutants Proceed Into the Lysosomal Degradatimentioning

confidence: 99%

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N‐glycosylation of the AMPA‐type glutamate receptor regulates cell surface expression and tetramer formation affecting channel function

Kandel

Yamamoto

Midorikawa

et al. 2018

Journal of Neurochemistry

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The AMPA-type glutamate receptor (AMPA-R) plays a primary role in principal excitatory synaptic transmission and many neuronal functions including synaptic plasticity that underlie learning and memory. N-glycosylation is one of the major post-translational modifications of membrane proteins, but its specific roles in neurons remain largely unknown. AMPA-R subunits are N-glycosylated at their extracellular domains during their biosynthesis in the lumen of the endoplasmic reticulum and Golgi system. Six N-glycosylation sites are presumed to exist in the extracellular domain of GluA1, which is a member of the AMPA-R subunits. We observed that the intracellular trafficking and cell surface expression were strongly suppressed in the GluA1 mutants lacking N-glycans at N63/N363 in HEK293T cells. Multimer analysis using Blue Native-PAGE displayed the impaired tetramer formation in the glycosylation mutants (N63S and N363S), indicating that the mis-transport was caused by impaired tetramer formation. N63S and N363S mutants were primarily degraded via the lysosomal pathway. Flag-tagged N363S GluA1, but not N63S GluA1, expressed in primary cortical neuron cultures prepared from GluA1 knockout mice was observed to localize at the cell surface. Co-expression of GluA2 partially rescued tetramer formation and the cell surface expression of N363S GluA1 but not N63S GluA1, in HEK293T cells. Electrophysiological analysis also demonstrated functional heteromers of N363S GluA1 with GluA2. These data suggest that site-specific N-glycans on GluA1 subunit regulates tetramer formation, intracellular trafficking, and cell surface expression of AMPA-R. OPEN PRACTICES: Open Science: This manuscript was awarded with the Open Materials Badge. For more information see: https://cos.io/our-services/open-science-badges/.

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“…On the other hand, mono‐ or short ubiquitination are involved in the endocytosis of membrane proteins, followed by sorting to the lysosome (Su et al . ; Goo et al . ).…”

Section: N‐glycosylation Mutants Proceed Into the Lysosomal Degradatimentioning

confidence: 99%

Section: N-glycosylation Mutants Proceed Into the Lysosomal Degradatimentioning

confidence: 99%

N‐glycosylation of the AMPA‐type glutamate receptor regulates cell surface expression and tetramer formation affecting channel function

Kandel

Yamamoto

Midorikawa

et al. 2018

Journal of Neurochemistry

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“…Phosphorylation often regulates ubiquitination of proteins, thereby promoting their degradation [39–41]. Several lines of evidence indicate that regulation of cell surface GABA B receptors by CaMKIIβ is conveyed by MIB2-mediated Lys-63-linked ubiquitination of GABA B1 .…”

Section: Discussionmentioning

confidence: 99%

Ca2+/Calmodulin-Dependent Protein Kinase II (CaMKII) β-Dependent Phosphorylation of GABAB1 Triggers Lysosomal Degradation of GABAB Receptors via Mind Bomb-2 (MIB2)-Mediated Lys-63-Linked Ubiquitination

et al. 2018

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The G protein-coupled GABA B receptors, constituted from GABA B1 and GABA B2 subunits, are important regulators of neuronal excitability by mediating long-lasting inhibition. One factor that determines receptor availability and thereby the strength of inhibition is regulated protein degradation. GABA B receptors are constitutively internalized from the plasma membrane and are either recycled to the cell surface or degraded in lysosomes. Lys-63-linked ubiquitination mediated by the E3 ligase Mind bomb-2 (MIB2) is the signal that sorts GABA B receptors to lysosomes. However, it is unknown how Lys-63-linked ubiquitination and thereby lysosomal degradation of the receptors is regulated. Here, we show that Ca 2+ /calmodulin-dependent protein kinase II (CaMKII) promotes MIB2-mediated Lys-63-linked ubiquitination of GABA B receptors. We found that inhibition of CaMKII in cultured rat cortical neurons increased cell surface GABA B receptors, whereas overexpression of CaMKIIβ, but not CaMKIIα, decreased receptor levels. This effect was conveyed by Lys-63-linked ubiquitination of GABA B1 at multiple sites mediated by the E3 ligase MIB2. Inactivation of the CaMKII phosphorylation site on GABA B1 (Ser-867) strongly reduced Lys-63-linked ubiquitination of GABA B receptors and increased their cell surface expression, whereas the phosphomimetic mutant GABA B1 (S867D) exhibited strongly increased Lys-63-linked ubiquitination and reduced cell surface expression. Finally, triggering lysosomal degradation of GABA B receptors by sustained activation of glutamate receptors, a condition occurring in brain ischemia, was accompanied with a massive increase of GABA B1 (Ser-867) phosphorylation-dependent Lys-63-linked ubiquitination of GABA B receptors. These findings indicate that CaMKIIβ-dependent Lys-63-linked ubiquitination of GABA B1 at multiple sites controls sorting of GABA B receptors to lysosomes for degradation under physiological and pathological condition.

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“…We recently reported that FSK, the adenylyl cyclase stimulator often used for in vitro experiments to stimulate UT-A1 urea transport activity, can also promote UT-A1 ubiquitination and protein degradation, revealing the other side of UT-A1 phosphorylation by vasopressin or FSK (29,30). This observation has led to the hypothesis that phosphorylation could be required for UT-A1 ubiquitination.…”

Section: F76mentioning

confidence: 99%

Downregulation of urea transporter UT-A1 activity by 14-3-3 protein

et al. 2015

American Journal of Physiology-Renal Physiology

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Urea transporter (UT)-A1 in the kidney inner medulla plays a critical role in the urinary concentrating mechanism and thereby in the regulation of water balance. The 14-3-3 proteins are a family of seven isoforms. They are multifunctional regulatory proteins that mainly bind to phosphorylated serine/threonine residues in target proteins. In the present study, we found that all seven 14-3-3 isoforms were detected in the kidney inner medulla. However, only the 14-3-3 γ-isoform was specifically and highly associated with UT-A1, as demonstrated by a glutathione- S-transferase-14-3-3 pulldown assay. The cAMP/adenylyl cyclase stimulator forskolin significantly enhanced their binding. Coinjection of 14-3-3γ cRNA into oocytes resulted in a decrease of UT-A1 function. In addition, 14-3-3γ increased UT-A1 ubiquitination and protein degradation. 14-3-3γ can interact with both UT-A1 and mouse double minute 2, the E3 ubiquitin ligase for UT-A1. Thus, activation of cAMP/PKA increases 14-3-3γ interactions with UT-A1 and stimulates mouse double minute 2-mediated UT-A1 ubiquitination and degradation, thereby forming a novel regulatory mechanism of urea transport activity.

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Activation of the cAMP/PKA pathway induces UT-A1 urea transporter monoubiquitination and targets it for lysosomal degradation

Cited by 14 publications

References 37 publications

N‐glycosylation of the AMPA‐type glutamate receptor regulates cell surface expression and tetramer formation affecting channel function

N‐glycosylation of the AMPA‐type glutamate receptor regulates cell surface expression and tetramer formation affecting channel function

Ca2+/Calmodulin-Dependent Protein Kinase II (CaMKII) β-Dependent Phosphorylation of GABAB1 Triggers Lysosomal Degradation of GABAB Receptors via Mind Bomb-2 (MIB2)-Mediated Lys-63-Linked Ubiquitination

Downregulation of urea transporter UT-A1 activity by 14-3-3 protein

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