Signal peptide represses GluK1 surface and synaptic trafficking through binding to amino-terminal domain

Duan, Guifang; Ye, Yaxin; Xu, Sha; Tao, Wucheng; Zhao, S. P.; Jin, Tengchuan; Shi, Yun; Sheng, Nengyin

doi:10.1038/s41467-018-07403-7

Cited by 17 publications

(34 citation statements)

References 28 publications

(45 reference statements)

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“…Previous work shows that Neto1 speeds and Neto2 slows GluK1 desensitization in a recombinant system or in neurons (28,31). Similarly, Neto1 speeds and Neto2 slows GluK2 desensitization when overexpressed in hippocampal CA1 neurons (11).…”

Section: Ntd-cub1 Interactions Distinguish Neto1 and Neto2 On Desensitizationmentioning

confidence: 92%

“…Previously, we found the synaptic targeting property of GluK1 and GluK2 is differentially regulated by Netos in an NTD-dependent manner (11,28,29), indicating that NTDs of KARs might directly interact with Netos. To test this hypothesis, we expressed the NTD of GluK2 by introducing a stop codon at position 401; thus, the NTD (residues 31-400) will be synthesized through the secretory ER pathway under the guidance of the signal peptide.…”

Section: Netos Interact With Gluk2 Through Multiple Sitesmentioning

confidence: 94%

“…We previously reported that synaptic targeting of GluK1 in hippocampal CA1 neurons relies on Neto proteins, whereas that of GluK2 does not (11,28). The differential trafficking properties and dependence on Netos between GluK1 and GluK2 rely on their NTDs (11,29).…”

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confidence: 99%

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Neto proteins regulate gating of the kainate-type glutamate receptor GluK2 through two binding sites

Duan

Jia

et al. 2019

Journal of Biological Chemistry

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Edited by Roger J. ColbranThe neuropilin and tolloid-like (Neto) proteins Neto1 and Neto2 are auxiliary subunits of kainate-type glutamate receptors (KARs) that regulate KAR trafficking and gating. However, how Netos bind and regulate the biophysical functions of KARs remains unclear. Here, we found that the N-terminal domain (NTD) of glutamate receptor ionotropic kainate 2 (GluK2) binds the first complement C1r/C1s-Uegf-BMP (CUB) domain of Neto proteins (i.e. NTD-CUB1 interaction) and that the core of GluK2 (GluK2⌬NTD) binds Netos through domains other than CUB1s (core-Neto interaction). Using electrophysiological analysis in HEK293T cells, we examined the effects of these interactions on GluK2 gating, including deactivation, desensitization, and recovery from desensitization. We found that NTD deletion does not affect GluK2 fast gating kinetics, the desensitization, and the deactivation. We also observed that Neto1 and Neto2 differentially regulate GluK2 fast gating kinetics, which largely rely on the NTD-CUB1 interactions. NTD removal facilitated GluK2 recovery from desensitization, indicating that the NTD stabilizes the GluK2 desensitization state. Co-expression with Neto1 or Neto2 also accelerated GluK2 recovery from desensitization, which fully relied on the NTD-CUB1 interactions. Moreover, we demonstrate that the NTD-CUB1 interaction involves electric attraction between positively charged residues in the GluK2_NTD and negatively charged ones in the CUB1 domains. Neutralization of these charges eliminated the regulatory effects of the NTD-CUB1 interaction on GluK2 gating. We conclude that KARs bind Netos through at least two sites and that the NTD-CUB1 interaction critically regulates Neto-mediated GluK2 gating.

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Section: Ntd-cub1 Interactions Distinguish Neto1 and Neto2 On Desensitizationmentioning

confidence: 92%

Section: Netos Interact With Gluk2 Through Multiple Sitesmentioning

confidence: 94%

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Neto proteins regulate gating of the kainate-type glutamate receptor GluK2 through two binding sites

Duan

Jia

et al. 2019

Journal of Biological Chemistry

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“…The full-length coding sequences of Tmem63b (NM_198167.3), Adar1 (Adar, NM_019655. 3) and Adar2 (Adarb1, NM_001024837.2) were amplified from mouse brain cDNAs, using specific primers (Table S4), by PrimeSTAR® HS DNA Polymerase (TaKaRa, R01A) and subcloned into pCAGGS vectors by Ligation-Free Cloning Kit (abm, E001) (50). For measuring the cytoplasmic calcium concentration, the free calcium indicator GCaMP6f was fused to the C-terminal of Tmem63b through a P2A linker (Tmem63b-P2A-GCaMP6f), lead to the separate expression of Tmem63b and GCaMP6f (27).…”

Section: Cloning Of Tmem63b Adar1 and Adar2 From Mouse Brainsmentioning

confidence: 99%

Distant coupling between RNA editing and alternative splicing of the osmosensitive cation channel Tmem63b

Zang

Shi

et al. 2020

Journal of Biological Chemistry

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Post-transcriptional modifications of pre-mRNAs expand the diversity of proteomes in higher eukaryotes. In brain, these modifications diversify the functional output of many critical neuronal signal molecules. In this study, we identified a brain-specific A-to-I RNA editing that changed glutamine to arginine (Q/R) at exon 20 and an alternative splicing of exon 4 in Tmem63b, which encodes a ubiquitously expressed osmosensitive cation channel. The channel isoforms lacking exon 4 occurred in ~80% of Tmem63b mRNAs in the brain but were not detected in other tissues, suggesting a brain-specific splicing. We found that the Q/R editing was catalyzed by Adar2 (Adarb1) and required an editing site complementary sequence located in the proximal 5’ end of intron 20. Moreover, the Q/R editing was almost exclusively identified in the splicing isoform lacking exon 4, indicating a coupling between the editing and the splicing. Elimination of the Q/R editing in brain-specific Adar2 knockout mice did not affect the splicing efficiency of exon 4. Furthermore, transfection with the splicing isoform containing exon 4 suppressed the Q/R editing in primary cultured cerebellar granule neurons. Thus, our study revealed a coupling between an RNA editing and a distant alternative splicing in the Tmem63b pre-mRNA, in which the splicing plays a dominant role. Finally, physiological analysis showed that the splicing and the editing coordinately regulate Ca2+ permeability and osmosensitivity of channel proteins, which may contribute to their functions in brain.

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“…Importantly, recent studies suggest that amino acids could have beneficial and detrimental effects. For example, glutamate plays an important role in maintaining the normal signal transduction of nerve cells, which is beneficial to the synaptic plasticity of neurons and to the recovery of stroke (3). However, elevated levels of glutamate can trigger oxidative stress, inflammation, and endothelial damage (4).…”

Section: Introductionmentioning

confidence: 99%

Targeted Metabolomic Profiling Reveals Association Between Altered Amino Acids and Poor Functional Recovery After Stroke

et al. 2020

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Amino acids have been shown to be among the most important metabolites to be altered following stroke; however, they are a double-edged sword with regard to regulating hemostasis. In this study, we conducted a targeted metabolomic study to examine the association between serum levels of amino acids and functional recovery after stroke. Three hundred and fifty-one patients with stroke admitted to an acute rehabilitation hospital were screened, and 106 patients were selected based on inclusion and exclusion criteria. Recruited patients were stratified using Montebello Rehabilitation Factor Score (MRFS) efficiency. We selected the top (n = 20, 19%) and bottom (n = 20, 19%) of MRFS efficiency for metabolomic analysis. A total of 21 serum amino acids levels were measured using ultra high performance liquid chromatography and mass spectrometry. The normalized data were analyzed by multivariate approaches, and the selected potential biomarkers were combined in different combinations for prediction of stroke functional recovery. The results demonstrated that there were significant differences in leucine-isoleucine, proline, threonine, glutamic acid, and arginine levels between good and poor recovery groups. In the training (0.952) and test (0.835) sets, metabolite biomarker panels composed of proline, glutamic acid, and arginine had the highest sensitivity and specificity in distinguishing good recovery from poor. In particular, arginine was present in the top 10 combinations of the average area under the receiver operating characteristic curve (AUC) test set. Our findings suggest that amino acids related to energy metabolism and excitotoxicity may play an important role in functional recovery after stroke. Therefore, the level of serum arginine has predictive value for the recovery rate after stroke.

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Signal peptide represses GluK1 surface and synaptic trafficking through binding to amino-terminal domain

Cited by 17 publications

References 28 publications

Neto proteins regulate gating of the kainate-type glutamate receptor GluK2 through two binding sites

Neto proteins regulate gating of the kainate-type glutamate receptor GluK2 through two binding sites

Distant coupling between RNA editing and alternative splicing of the osmosensitive cation channel Tmem63b

Targeted Metabolomic Profiling Reveals Association Between Altered Amino Acids and Poor Functional Recovery After Stroke

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