Yoo, Jae Cheal Park, Nammi Lee, Boah Noshed, Abdullateef Lee, Young-Sun Kim, Tae Hwan Lee, Da Yong Kim, Ajung Hwang, Eun Mi Yi, Gwan-su Park, Jae-Yong 2024-01-20T01:03:04Z 2024-01-20T01:03:04Z 2021-09-04 2017-07-01 0014-4827 https://pubs.kist.re.kr/handle/201004/122549 Copinel (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, 14-3-3 gamma appears to directly interact with CPNE1. Between CPNE1 and 14-3-3 gamma, 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 14-3-3 gamma directly interacts with CPNE1. Our results also demonstrate that AKT phosphorylation, neurite outgrowth and expression of the neuronal marker protein are increased when 14-3-3 gamma 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 14-3-3 gamma, and in differentiation-related function of CPNE1. Moreover, mutation of Ser54 amino acids residue in CPNE1 effectively decreased association with 14-3-3 gamma and neuronal differentiation of HiB5 cells. Collectively, our findings indicate that 14-3-3 gamma regulates the differentiation ability of CPNE1 through the binding with C2A domain of CPNE1 in HiB5 cells. English ELSEVIER INC PROTEINS BINDING EXPRESSION BRAIN IDENTIFICATION COFILIN FAMILY DOMAIN 14-3-3 gamma regulates Copinel-mediated neuronal differentiation in HiB5 hippocampal progenitor cells Article 10.1016/j.yexcr.2017.04.015 1 EXPERIMENTAL CELL RESEARCH, v.356, no.1, pp.85 - 92 EXPERIMENTAL CELL RESEARCH 356 1 85 92 scie scopus 000402776700010 2-s2.0-85017426563 Oncology Cell Biology Oncology Cell Biology Article PROTEINS BINDING EXPRESSION BRAIN IDENTIFICATION COFILIN FAMILY DOMAIN Copinel 14-3-3 gamma Protein-protein interaction C2A domain HiB5 cell