Direct interaction with 14-3-3γ promotes surface expression of Best1 channel in astrocyte

Soo Jin Oh, Junsung Woo, Young Sun Lee, Minhee Cho, Eunju Kim, Nam Chul Cho, Jae-Yong Park, Ae Nim Pae, Changjoon Lee, Eun Mi Hwang

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background: Bestrophin-1 (Best1) is a calcium-activated anion channel (CAAC) that is expressed broadly in mammalian tissues including the brain. We have previously reported that Best1 is expressed in hippocampal astrocytes at the distal peri-synaptic regions, called microdomains, right next to synaptic junctions, and that it disappears from the microdomains in Alzheimer's disease mouse model. Although Best1 appears to be dynamically regulated, the mechanism of its regulation and modulation is poorly understood. It has been reported that a regulatory protein, 14-3-3 affects the surface expression of numerous membrane proteins in mammalian cells. Methods: The protein-protein interaction between Best1 and 14-3-3γ was confirmed by yeast-two hybrid assay and BiFC method. The effect of 14-3-3γ on Best1-mediated current was measured by whole-cell patch clamp technique. Results: We identified 14-3-3γ as novel binding partner of Best1 in astrocytes: among 7 isoforms of 14-3-3 protein, only 14-3-3γ was found to bind specifically. We determined a binding domain on the C-terminus of Best1 which is critical for an interaction with 14-3-3γ. We also revealed that interaction between Best1 and 14-3-3γ was mediated by phosphorylation of S358 in the C-terminus of Best1. We confirmed that surface expression of Best1 and Best1-mediated whole-cell current were significantly decreased after a gene-silencingof 14-3-3γ without a significant change in total Best1 expression in cultured astrocytes. Furthermore, we discovered that 14-3-3γ-shRNA reduced Best1-mediated glutamate release from hippocampal astrocyte by recording a PAR1 receptor-induced NMDA receptor-mediated current from CA1 pyramidal neurons in hippocampal slices injected with adenovirus carrying 14-3-3γ-shRNA. Finally, through a structural modeling, we found critical amino acid residues containing S358 of Best1 exhibiting binding affinities to 14-3-3γ. Conclusions: 14-3-3γ promotes surface expression of Best1 channel in astrocytes through direct interaction.

Original languageEnglish
Article number51
JournalMolecular Brain
Volume10
Issue number1
DOIs
Publication statusPublished - 2017 Nov 9

Fingerprint

14-3-3 Proteins
Astrocytes
Small Interfering RNA
PAR-1 Receptor
Two-Hybrid System Techniques
Pyramidal Cells
Patch-Clamp Techniques
N-Methyl-D-Aspartate Receptors
Adenoviridae
Anions
Glutamic Acid
Alzheimer Disease
Protein Isoforms
Membrane Proteins
Proteins
Phosphorylation
Calcium
Amino Acids
Brain
Genes

Keywords

  • 14-3-3γ
  • Astrocyte
  • Bestrophin-1
  • Glutamate
  • Surface expression

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

Cite this

Oh, S. J., Woo, J., Lee, Y. S., Cho, M., Kim, E., Cho, N. C., ... Hwang, E. M. (2017). Direct interaction with 14-3-3γ promotes surface expression of Best1 channel in astrocyte. Molecular Brain, 10(1), [51]. https://doi.org/10.1186/s13041-017-0331-x

Direct interaction with 14-3-3γ promotes surface expression of Best1 channel in astrocyte. / Oh, Soo Jin; Woo, Junsung; Lee, Young Sun; Cho, Minhee; Kim, Eunju; Cho, Nam Chul; Park, Jae-Yong; Pae, Ae Nim; Lee, Changjoon; Hwang, Eun Mi.

In: Molecular Brain, Vol. 10, No. 1, 51, 09.11.2017.

Research output: Contribution to journalArticle

Oh, SJ, Woo, J, Lee, YS, Cho, M, Kim, E, Cho, NC, Park, J-Y, Pae, AN, Lee, C & Hwang, EM 2017, 'Direct interaction with 14-3-3γ promotes surface expression of Best1 channel in astrocyte', Molecular Brain, vol. 10, no. 1, 51. https://doi.org/10.1186/s13041-017-0331-x
Oh, Soo Jin ; Woo, Junsung ; Lee, Young Sun ; Cho, Minhee ; Kim, Eunju ; Cho, Nam Chul ; Park, Jae-Yong ; Pae, Ae Nim ; Lee, Changjoon ; Hwang, Eun Mi. / Direct interaction with 14-3-3γ promotes surface expression of Best1 channel in astrocyte. In: Molecular Brain. 2017 ; Vol. 10, No. 1.
@article{6c232019defe46d5865216ab77c4bdb5,
title = "Direct interaction with 14-3-3γ promotes surface expression of Best1 channel in astrocyte",
abstract = "Background: Bestrophin-1 (Best1) is a calcium-activated anion channel (CAAC) that is expressed broadly in mammalian tissues including the brain. We have previously reported that Best1 is expressed in hippocampal astrocytes at the distal peri-synaptic regions, called microdomains, right next to synaptic junctions, and that it disappears from the microdomains in Alzheimer's disease mouse model. Although Best1 appears to be dynamically regulated, the mechanism of its regulation and modulation is poorly understood. It has been reported that a regulatory protein, 14-3-3 affects the surface expression of numerous membrane proteins in mammalian cells. Methods: The protein-protein interaction between Best1 and 14-3-3γ was confirmed by yeast-two hybrid assay and BiFC method. The effect of 14-3-3γ on Best1-mediated current was measured by whole-cell patch clamp technique. Results: We identified 14-3-3γ as novel binding partner of Best1 in astrocytes: among 7 isoforms of 14-3-3 protein, only 14-3-3γ was found to bind specifically. We determined a binding domain on the C-terminus of Best1 which is critical for an interaction with 14-3-3γ. We also revealed that interaction between Best1 and 14-3-3γ was mediated by phosphorylation of S358 in the C-terminus of Best1. We confirmed that surface expression of Best1 and Best1-mediated whole-cell current were significantly decreased after a gene-silencingof 14-3-3γ without a significant change in total Best1 expression in cultured astrocytes. Furthermore, we discovered that 14-3-3γ-shRNA reduced Best1-mediated glutamate release from hippocampal astrocyte by recording a PAR1 receptor-induced NMDA receptor-mediated current from CA1 pyramidal neurons in hippocampal slices injected with adenovirus carrying 14-3-3γ-shRNA. Finally, through a structural modeling, we found critical amino acid residues containing S358 of Best1 exhibiting binding affinities to 14-3-3γ. Conclusions: 14-3-3γ promotes surface expression of Best1 channel in astrocytes through direct interaction.",
keywords = "14-3-3γ, Astrocyte, Bestrophin-1, Glutamate, Surface expression",
author = "Oh, {Soo Jin} and Junsung Woo and Lee, {Young Sun} and Minhee Cho and Eunju Kim and Cho, {Nam Chul} and Jae-Yong Park and Pae, {Ae Nim} and Changjoon Lee and Hwang, {Eun Mi}",
year = "2017",
month = "11",
day = "9",
doi = "10.1186/s13041-017-0331-x",
language = "English",
volume = "10",
journal = "Molecular Brain",
issn = "1756-6606",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Direct interaction with 14-3-3γ promotes surface expression of Best1 channel in astrocyte

AU - Oh, Soo Jin

AU - Woo, Junsung

AU - Lee, Young Sun

AU - Cho, Minhee

AU - Kim, Eunju

AU - Cho, Nam Chul

AU - Park, Jae-Yong

AU - Pae, Ae Nim

AU - Lee, Changjoon

AU - Hwang, Eun Mi

PY - 2017/11/9

Y1 - 2017/11/9

N2 - Background: Bestrophin-1 (Best1) is a calcium-activated anion channel (CAAC) that is expressed broadly in mammalian tissues including the brain. We have previously reported that Best1 is expressed in hippocampal astrocytes at the distal peri-synaptic regions, called microdomains, right next to synaptic junctions, and that it disappears from the microdomains in Alzheimer's disease mouse model. Although Best1 appears to be dynamically regulated, the mechanism of its regulation and modulation is poorly understood. It has been reported that a regulatory protein, 14-3-3 affects the surface expression of numerous membrane proteins in mammalian cells. Methods: The protein-protein interaction between Best1 and 14-3-3γ was confirmed by yeast-two hybrid assay and BiFC method. The effect of 14-3-3γ on Best1-mediated current was measured by whole-cell patch clamp technique. Results: We identified 14-3-3γ as novel binding partner of Best1 in astrocytes: among 7 isoforms of 14-3-3 protein, only 14-3-3γ was found to bind specifically. We determined a binding domain on the C-terminus of Best1 which is critical for an interaction with 14-3-3γ. We also revealed that interaction between Best1 and 14-3-3γ was mediated by phosphorylation of S358 in the C-terminus of Best1. We confirmed that surface expression of Best1 and Best1-mediated whole-cell current were significantly decreased after a gene-silencingof 14-3-3γ without a significant change in total Best1 expression in cultured astrocytes. Furthermore, we discovered that 14-3-3γ-shRNA reduced Best1-mediated glutamate release from hippocampal astrocyte by recording a PAR1 receptor-induced NMDA receptor-mediated current from CA1 pyramidal neurons in hippocampal slices injected with adenovirus carrying 14-3-3γ-shRNA. Finally, through a structural modeling, we found critical amino acid residues containing S358 of Best1 exhibiting binding affinities to 14-3-3γ. Conclusions: 14-3-3γ promotes surface expression of Best1 channel in astrocytes through direct interaction.

AB - Background: Bestrophin-1 (Best1) is a calcium-activated anion channel (CAAC) that is expressed broadly in mammalian tissues including the brain. We have previously reported that Best1 is expressed in hippocampal astrocytes at the distal peri-synaptic regions, called microdomains, right next to synaptic junctions, and that it disappears from the microdomains in Alzheimer's disease mouse model. Although Best1 appears to be dynamically regulated, the mechanism of its regulation and modulation is poorly understood. It has been reported that a regulatory protein, 14-3-3 affects the surface expression of numerous membrane proteins in mammalian cells. Methods: The protein-protein interaction between Best1 and 14-3-3γ was confirmed by yeast-two hybrid assay and BiFC method. The effect of 14-3-3γ on Best1-mediated current was measured by whole-cell patch clamp technique. Results: We identified 14-3-3γ as novel binding partner of Best1 in astrocytes: among 7 isoforms of 14-3-3 protein, only 14-3-3γ was found to bind specifically. We determined a binding domain on the C-terminus of Best1 which is critical for an interaction with 14-3-3γ. We also revealed that interaction between Best1 and 14-3-3γ was mediated by phosphorylation of S358 in the C-terminus of Best1. We confirmed that surface expression of Best1 and Best1-mediated whole-cell current were significantly decreased after a gene-silencingof 14-3-3γ without a significant change in total Best1 expression in cultured astrocytes. Furthermore, we discovered that 14-3-3γ-shRNA reduced Best1-mediated glutamate release from hippocampal astrocyte by recording a PAR1 receptor-induced NMDA receptor-mediated current from CA1 pyramidal neurons in hippocampal slices injected with adenovirus carrying 14-3-3γ-shRNA. Finally, through a structural modeling, we found critical amino acid residues containing S358 of Best1 exhibiting binding affinities to 14-3-3γ. Conclusions: 14-3-3γ promotes surface expression of Best1 channel in astrocytes through direct interaction.

KW - 14-3-3γ

KW - Astrocyte

KW - Bestrophin-1

KW - Glutamate

KW - Surface expression

UR - http://www.scopus.com/inward/record.url?scp=85033595831&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85033595831&partnerID=8YFLogxK

U2 - 10.1186/s13041-017-0331-x

DO - 10.1186/s13041-017-0331-x

M3 - Article

C2 - 29121962

AN - SCOPUS:85033595831

VL - 10

JO - Molecular Brain

JF - Molecular Brain

SN - 1756-6606

IS - 1

M1 - 51

ER -