14-3-3γ regulates Copine1-mediated neuronal differentiation in HiB5 hippocampal progenitor cells

Jae Cheal Yoo, Nammi Park, Boah Lee, Abdullateef Nashed, Young Sun Lee, Tae Hwan Kim, Da Yong Lee, Ajung Kim, Eun Mi Hwang, Gwan su Yi, Jae-Yong Park

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Copine1 (CPNE1), known as a calcium-dependent membrane-binding protein, has tandem C2 domains and an A domain. We previously demonstrated that CPNE1 directly induces neuronal differentiation via Protein kinase B (AKT) phosphorylation in the hippocampal progenitor cell line, HiB5. To better understand its cellular function, we carried out a yeast two-hybrid screening to find CPNE1 binding partners. Among the identified proteins, 14-3-3γ appears to directly interact with CPNE1. Between CPNE1 and 14-3-3γ, the physical interaction as well as the specific binding regions of CPNE1 was confirmed in vitro and in vivo. Furthermore, among the seven 14-3-3 isotypes, only 14-3-3γ directly interacts with CPNE1. Our results also demonstrate that AKT phosphorylation, neurite outgrowth and expression of the neuronal marker protein are increased when 14-3-3γ is overexpressed in CPNE1 high expressed HiB5 cells. Furthermore, the neighboring Ser54 amino acids residue of C2A domain in CPNE1 has an important role in binding with 14-3-3γ, and in differentiation-related function of CPNE1. Moreover, mutation of Ser54 amino acids residue in CPNE1 effectively decreased association with 14-3-3γ and neuronal differentiation of HiB5 cells. Collectively, our findings indicate that 14-3-3γ regulates the differentiation ability of CPNE1 through the binding with C2A domain of CPNE1 in HiB5 cells.

Original languageEnglish
Pages (from-to)85-92
Number of pages8
JournalExperimental Cell Research
Volume356
Issue number1
DOIs
Publication statusPublished - 2017 Jul 1

Fingerprint

Stem Cells
Phosphorylation
14-3-3 Proteins
Amino Acids
Proto-Oncogene Proteins c-akt
Cell Differentiation
Carrier Proteins
Membrane Proteins
Yeasts
Calcium
Cell Line
Mutation
Proteins
Neuronal Outgrowth
In Vitro Techniques
C2 Domains

Keywords

  • 14-3-3γ
  • C2A domain
  • Copine1
  • HiB5 cell
  • Protein-protein interaction

ASJC Scopus subject areas

  • Cell Biology

Cite this

14-3-3γ regulates Copine1-mediated neuronal differentiation in HiB5 hippocampal progenitor cells. / Cheal Yoo, Jae; Park, Nammi; Lee, Boah; Nashed, Abdullateef; Lee, Young Sun; Hwan Kim, Tae; Yong Lee, Da; Kim, Ajung; Mi Hwang, Eun; Yi, Gwan su; Park, Jae-Yong.

In: Experimental Cell Research, Vol. 356, No. 1, 01.07.2017, p. 85-92.

Research output: Contribution to journalArticle

Cheal Yoo, J, Park, N, Lee, B, Nashed, A, Lee, YS, Hwan Kim, T, Yong Lee, D, Kim, A, Mi Hwang, E, Yi, GS & Park, J-Y 2017, '14-3-3γ regulates Copine1-mediated neuronal differentiation in HiB5 hippocampal progenitor cells', Experimental Cell Research, vol. 356, no. 1, pp. 85-92. https://doi.org/10.1016/j.yexcr.2017.04.015
Cheal Yoo, Jae ; Park, Nammi ; Lee, Boah ; Nashed, Abdullateef ; Lee, Young Sun ; Hwan Kim, Tae ; Yong Lee, Da ; Kim, Ajung ; Mi Hwang, Eun ; Yi, Gwan su ; Park, Jae-Yong. / 14-3-3γ regulates Copine1-mediated neuronal differentiation in HiB5 hippocampal progenitor cells. In: Experimental Cell Research. 2017 ; Vol. 356, No. 1. pp. 85-92.
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AU - Nashed, Abdullateef

AU - Lee, Young Sun

AU - Hwan Kim, Tae

AU - Yong Lee, Da

AU - Kim, Ajung

AU - Mi Hwang, Eun

AU - Yi, Gwan su

AU - Park, Jae-Yong

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AB - Copine1 (CPNE1), known as a calcium-dependent membrane-binding protein, has tandem C2 domains and an A domain. We previously demonstrated that CPNE1 directly induces neuronal differentiation via Protein kinase B (AKT) phosphorylation in the hippocampal progenitor cell line, HiB5. To better understand its cellular function, we carried out a yeast two-hybrid screening to find CPNE1 binding partners. Among the identified proteins, 14-3-3γ appears to directly interact with CPNE1. Between CPNE1 and 14-3-3γ, the physical interaction as well as the specific binding regions of CPNE1 was confirmed in vitro and in vivo. Furthermore, among the seven 14-3-3 isotypes, only 14-3-3γ directly interacts with CPNE1. Our results also demonstrate that AKT phosphorylation, neurite outgrowth and expression of the neuronal marker protein are increased when 14-3-3γ is overexpressed in CPNE1 high expressed HiB5 cells. Furthermore, the neighboring Ser54 amino acids residue of C2A domain in CPNE1 has an important role in binding with 14-3-3γ, and in differentiation-related function of CPNE1. Moreover, mutation of Ser54 amino acids residue in CPNE1 effectively decreased association with 14-3-3γ and neuronal differentiation of HiB5 cells. Collectively, our findings indicate that 14-3-3γ regulates the differentiation ability of CPNE1 through the binding with C2A domain of CPNE1 in HiB5 cells.

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