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RIBEYE B结构域对于RIBEYE A结构域的稳定性和突触 ribbons 的组装至关重要。

RIBEYE B-Domain Is Essential for RIBEYE A-Domain Stability and Assembly of Synaptic Ribbons.

作者信息

Shankhwar Soni, Schwarz Karin, Katiyar Rashmi, Jung Martin, Maxeiner Stephan, Südhof Thomas C, Schmitz Frank

机构信息

Institute of Anatomy and Cell Biology, Saarland University, Medical School, Homburg, Germany.

Institute of Medical Biochemistry and Molecular Biology, Saarland University, Medical School, Homburg, Germany.

出版信息

Front Mol Neurosci. 2022 Jan 28;15:838311. doi: 10.3389/fnmol.2022.838311. eCollection 2022.

DOI:10.3389/fnmol.2022.838311
PMID:35153673
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8831697/
Abstract

Synaptic ribbons are presynaptic specializations that define eponymous ribbon synapses. Synaptic ribbons are largely composed of RIBEYE, a protein containing an N-terminal A-domain and a carboxyterminal B-domain that is identical with CtBP2, a NAD(H)-binding transcriptional co-repressor. Previously we showed that synaptic ribbons are completely absent in RIBEYE knockout mice in which the RIBEYE A-domain-encoding exon had been deleted, but CtBP2 is still made, demonstrating that the A-domain is required for synaptic ribbon assembly. In the present study, we asked whether the RIBEYE B-domain also has an essential role in the assembly of synaptic ribbons. For this purpose, we made use of RIBEYE knockin mice in which the RIBEYE B-domain was replaced by a fluorescent protein domain, whereas the RIBEYE A-domain was retained unchanged. We found that replacing the RIBEYE B-domain with a fluorescent protein module destabilizes the resulting hybrid protein and causes a complete loss of synaptic ribbons. Our results thus demonstrate an essential role of the RIBEYE B-domain in enabling RIBEYE assembly into synaptic ribbons, reinforcing the notion that RIBEYE is the central organizer of synaptic ribbons.

摘要

突触带是定义了同名带状突触的突触前特化结构。突触带主要由RIBEYE组成，RIBEYE是一种蛋白质，包含一个N端A结构域和一个与CtBP2相同的C端B结构域，CtBP2是一种与NAD(H)结合的转录共抑制因子。此前我们发现，在敲除了编码RIBEYE A结构域的外显子的RIBEYE基因敲除小鼠中，突触带完全缺失，但仍能产生CtBP2，这表明A结构域是突触带组装所必需的。在本研究中，我们探究了RIBEYE B结构域在突触带组装中是否也具有重要作用。为此，我们利用了RIBEYE基因敲入小鼠，其中RIBEYE B结构域被一个荧光蛋白结构域取代，而RIBEYE A结构域保持不变。我们发现，用荧光蛋白模块取代RIBEYE B结构域会使产生的杂合蛋白不稳定，并导致突触带完全丧失。因此，我们的结果证明了RIBEYE B结构域在使RIBEYE组装成突触带方面的重要作用，强化了RIBEYE是突触带核心组织者的观点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/319f/8831697/12c9b27ce71c/fnmol-15-838311-g0013.jpg
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