Abstract
Excitatory neurotransmission relies on the precise targeting of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors to the neuronal plasma membrane. Activity-dependent ubiquitination of AMPA receptor (AMPAR) subunits sorts internalised receptors to late endosomes for degradation, which ultimately determines the number of AMPARs on neuronal membrane. Our recent study has demonstrated a functional cross-talk between the phosphorylation and ubiquitination of the GluA1 subunit in mammalian central neurons. However, the existence of such a cross modulation for the GluA2 subunit remains unknown. Here, we have shown that bicuculline induced GluA2 ubiquitination on the same lysine residues (Lys-870 and Lys-882) in the C-terminal as those elicited by the AMPA treatment. Interestingly, bicuculline-induced ubiquitination was markedly enhanced by the phospho-mimetic GluA2 S880E mutant. Pharmacological activation of protein kinase C (PKC) by phorbol ester, which mediates the phosphorylation of GluA2 at Ser-880, augmented bicuculline-induced ubiquitination of GluA2 in cultured neurons. This effect was specific for the GluA2 subunit because phorbol ester did not alter the level of GluA1 ubiquitination. However, phorbol ester-induced enhancement of GluA2 ubiquitination did not require Ser-880 phosphorylation. This suggests that pseudo-phosphorylation of Ser-880 is sufficient but is not necessary for the augmentation of bicuculline-induced GluA2 ubiquitination. Collectively, these data provide the first demonstration of subunit-specific modulation of AMPAR ubiquitination by the PKC-dependent signalling pathway in mammalian central neurons.
Similar content being viewed by others
Abbreviations
- AMPA:
-
α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid
- AMPARs:
-
AMPA receptors
- DIV:
-
Days in vitro
- DMSO:
-
Dimethyl sulfoxide
- GRIP1:
-
Glutamate receptor-interacting protein 1
- L-VGCCs:
-
L-type voltage-gated calcium channels
- LTD:
-
Long-term depression
- mGluRs:
-
Metabotropic glutamate receptors
- NEM:
-
N-Ethylmaleimide
- NMDA:
-
N-Methyl-d-aspartate
- NMDARs:
-
NMDA receptors
- PDZ:
-
Postsynaptic density 95/disc large/zona occludens-1
- PICK1:
-
Protein interacting with C-kinase 1
- PKA:
-
Protein kinase A
- PKC:
-
Protein kinase C
- PMA:
-
Phorbol 12-myristate 13-acetate
- RNF:
-
Ring finger protein
- WT:
-
Wild-type
Referencess
Anggono V, Huganir RL (2012) Regulation of AMPA receptor trafficking and synaptic plasticity. Curr Opin Neurobiol 22(3):461–469. https://doi.org/10.1016/j.conb.2011.12.006
Anggono V, Koc-Schmitz Y, Widagdo J, Kormann J, Quan A, Chen CM, Robinson PJ, Choi SY, Linden DJ, Plomann M, Huganir RL (2013) PICK1 interacts with PACSIN to regulate AMPA receptor internalization and cerebellar long-term depression. Proc Natl Acad Sci USA 110(34):13976–13981. https://doi.org/10.1073/pnas.1312467110
Bissen D, Foss F, Acker-Palmer A (2019) AMPA receptors and their minions: auxiliary proteins in AMPA receptor trafficking. Cell Mol Life Sci 76(11):2133–2169. https://doi.org/10.1007/s00018-019-03068-7
Bocock JP, Carmicle S, Madamba E, Erickson AH (2010) Nuclear targeting of an endosomal E3 ubiquitin ligase. Traffic 11(6):756–766. https://doi.org/10.1111/j.1600-0854.2010.01060.x
Boehm J, Kang MG, Johnson RC, Esteban J, Huganir RL, Malinow R (2006) Synaptic incorporation of AMPA receptors during LTP is controlled by a PKC phosphorylation site on GluR1. Neuron 51(2):213–225. https://doi.org/10.1016/j.neuron.2006.06.013
Chung HJ, Xia J, Scannevin RH, Zhang X, Huganir RL (2000) Phosphorylation of the AMPA receptor subunit GluR2 differentially regulates its interaction with PDZ domain-containing proteins. J Neurosci 20(19):7258–7267. https://doi.org/10.1523/JNEUROSCI.20-19-07258.2000
Chung HJ, Steinberg JP, Huganir RL, Linden DJ (2003) Requirement of AMPA receptor GluR2 phosphorylation for cerebellar long-term depression. Science 300(5626):1751–1755. https://doi.org/10.1126/science.1082915
Diering GH, Huganir RL (2018) The AMPA receptor code of synaptic plasticity. Neuron 100(2):314–329. https://doi.org/10.1016/j.neuron.2018.10.018
Dong H, O'Brien RJ, Fung ET, Lanahan AA, Worley PF, Huganir RL (1997) GRIP: a synaptic PDZ domain-containing protein that interacts with AMPA receptors. Nature 386(6622):279–284. https://doi.org/10.1038/386279a0
Fiuza M, Rostosky CM, Parkinson GT, Bygrave AM, Halemani N, Baptista M, Milosevic I, Hanley JG (2017) PICK1 regulates AMPA receptor endocytosis via direct interactions with AP2 α-appendage and dynamin. J Cell Biol 216(10):3323–3338. https://doi.org/10.1083/jcb.201701034
Goo MS, Scudder SL, Patrick GN (2015) Ubiquitin-dependent trafficking and turnover of ionotropic glutamate receptors. Front Mol Neurosci 8:60. https://doi.org/10.3389/fnmol.2015.00060
Guntupalli S, Jang SE, Zhu T, Huganir RL, Widagdo J, Anggono V (2017) GluA1 subunit ubiquitination mediates amyloid-β-induced loss of surface α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors. J Biol Chem 292(20):8186–8194. https://doi.org/10.1074/jbc.M116.774554
Hershko A, Ciechanover A (1998) The ubiquitin system. Annu Rev Biochem 67:425–479. https://doi.org/10.1146/annurev.biochem.67.1.425
Huganir RL, Nicoll RA (2013) AMPARs and synaptic plasticity: the last 25 years. Neuron 80(3):704–717. https://doi.org/10.1016/j.neuron.2013.10.025
Kemp BE, Pearson RB (1990) Protein kinase recognition sequence motifs. Trends Biochem Sci 15(9):342–346. https://doi.org/10.1016/0968-0004(90)90073-k
Lin DT, Huganir RL (2007) PICK1 and phosphorylation of the glutamate receptor 2 (GluR2) AMPA receptor subunit regulates GluR2 recycling after NMDA receptor-induced internalization. J Neurosci 27(50):13903–13908. https://doi.org/10.1523/JNEUROSCI.1750-07.2007
Lin DT, Makino Y, Sharma K, Hayashi T, Neve R, Takamiya K, Huganir RL (2009) Regulation of AMPA receptor extrasynaptic insertion by 4.1N, phosphorylation and palmitoylation. Nat Neurosci 12(7):879–887. https://doi.org/10.1038/nn.2351
Lin A, Hou Q, Jarzylo L, Amato S, Gilbert J, Shang F, Man HY (2011) Nedd4-mediated AMPA receptor ubiquitination regulates receptor turnover and trafficking. J Neurochem 119(1):27–39. https://doi.org/10.1111/j.1471-4159.2011.07221.x
Lu W, Shi Y, Jackson AC, Bjorgan K, During MJ, Sprengel R, Seeburg PH, Nicoll RA (2009) Subunit composition of synaptic AMPA receptors revealed by a single-cell genetic approach. Neuron 62(2):254–268. https://doi.org/10.1016/j.neuron.2009.02.027
Lussier MP, Nasu-Nishimura Y, Roche KW (2011) Activity-dependent ubiquitination of the AMPA receptor subunit GluA2. J Neurosci 31(8):3077–3081. https://doi.org/10.1523/JNEUROSCI.5944-10.2011
Lussier MP, Herring BE, Nasu-Nishimura Y, Neutzner A, Karbowski M, Youle RJ, Nicoll RA, Roche KW (2012) Ubiquitin ligase RNF167 regulates AMPA receptor-mediated synaptic transmission. Proc Natl Acad Sci USA 109(47):19426–19431. https://doi.org/10.1073/pnas.1217477109
Lussier MP, Sanz-Clemente A, Roche KW (2015) Dynamic regulation of N-methyl-D-aspartate (NMDA) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors by posttranslational modifications. J Biol Chem 290(48):28596–28603. https://doi.org/10.1074/jbc.R115.652750
Mabb AM, Ehlers MD (2010) Ubiquitination in postsynaptic function and plasticity. Annu Rev Cell Dev Biol 26:179–210. https://doi.org/10.1146/annurev-cellbio-100109-104129
Madsen KL, Thorsen TS, Rahbek-Clemmensen T, Eriksen J, Gether U (2012) Protein interacting with C kinase 1 (PICK1) reduces reinsertion rates of interaction partners sorted to Rab11-dependent slow recycling pathway. J Biol Chem 287(15):12293–12308. https://doi.org/10.1074/jbc.M111.294702
Rocca DL, Martin S, Jenkins EL, Hanley JG (2008) Inhibition of Arp2/3-mediated actin polymerization by PICK1 regulates neuronal morphology and AMPA receptor endocytosis. Nat Cell Biol 10(3):259–271. https://doi.org/10.1038/ncb1688
Schwarz LA, Hall BJ, Patrick GN (2010) Activity-dependent ubiquitination of GluA1 mediates a distinct AMPA receptor endocytosis and sorting pathway. J Neurosci 30(49):16718–16729. https://doi.org/10.1523/JNEUROSCI.3686-10.2010
Seidenman KJ, Steinberg JP, Huganir R, Malinow R (2003) Glutamate receptor subunit 2 Serine 880 phosphorylation modulates synaptic transmission and mediates plasticity in CA1 pyramidal cells. J Neurosci 23(27):9220–9228. https://doi.org/10.1523/JNEUROSCI.23-27-09220.2003
Selvakumar B, Jenkins MA, Hussain NK, Huganir RL, Traynelis SF, Snyder SH (2013) S-Nitrosylation of AMPA receptor GluA1 regulates phosphorylation, single-channel conductance, and endocytosis. Proc Natl Acad Sci USA 110(3):1077–1082. https://doi.org/10.1073/pnas.1221295110
Steinberg JP, Takamiya K, Shen Y, Xia J, Rubio ME, Yu S, Jin W, Thomas GM, Linden DJ, Huganir RL (2006) Targeted in vivo mutations of the AMPA receptor subunit GluR2 and its interacting protein PICK1 eliminate cerebellar long-term depression. Neuron 49(6):845–860. https://doi.org/10.1016/j.neuron.2006.02.025
Tan MC, Widagdo J, Chau YQ, Zhu T, Wong JJ, Cheung A, Anggono V (2017) The activity-induced long non-coding RNA Meg3 modulates AMPA receptor surface expression in primary cortical neurons. Front Cell Neurosci 11:124. https://doi.org/10.3389/fncel.2017.00124
Widagdo J, Chai YJ, Ridder MC, Chau YQ, Johnson RC, Sah P, Huganir RL, Anggono V (2015) Activity-dependent ubiquitination of GluA1 and GluA2 regulates AMPA receptor intracellular sorting and degradation. Cell Rep 10:783–795. https://doi.org/10.1016/j.celrep.2015.01.015
Widagdo J, Fang H, Jang SE, Anggono V (2016) PACSIN1 regulates the dynamics of AMPA receptor trafficking. Sci Rep 6:31070. https://doi.org/10.1038/srep31070
Widagdo J, Guntupalli S, Jang SE, Anggono V (2017) Regulation of AMPA receptor trafficking by protein ubiquitination. Front Mol Neurosci 10:347. https://doi.org/10.3389/fnmol.2017.00347
Xia J, Zhang X, Staudinger J, Huganir RL (1999) Clustering of AMPA receptors by the synaptic PDZ domain-containing protein PICK1. Neuron 22(1):179–187. https://doi.org/10.1016/S0896-6273(00)80689-3
Acknowledgments
This work was supported in part by grants from the Australian National Health and Medical Research Council (GNT1138452), the Alzheimer’s Australia Dementia Research Foundation (DPG14-57), and the Clem Jones Centre for Ageing Dementia Research (to VA). JW was supported by an Australian Research Council DECRA Fellowship (DE170100112). SG was a recipient of University of Queensland International Scholarship.
Author information
Authors and Affiliations
Contributions
VA conceived and supervised the project. VA and JW designed experiments. JW, JWK and SG performed research. RLH contributed molecular reagents. JW, JWK and VA wrote the manuscript. All authors analysed the results and approved the final version of the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
All authors declare that they have no competing financial interests.
Ethical Approval
All animal handling procedures were carried out in accordance to the Australian code for the care and use of animals for scientific purposes by the National Health and Medical Research Council and were approved by the University of Queensland Animal Ethics Committee (AEC approval number QBI/047/18).
Informed Consent
This article does not contain any studies with human participants performed by any of the authors. All authors have read and agreed with the submission of the manuscript.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Widagdo, J., Kerk, J.W., Guntupalli, S. et al. Subunit-Specific Augmentation of AMPA Receptor Ubiquitination by Phorbol Ester. Cell Mol Neurobiol 40, 1213–1222 (2020). https://doi.org/10.1007/s10571-020-00809-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10571-020-00809-2