Targeting of δ-catenin to postsynaptic sites through interaction with the Shank3 N-terminus.
Targeting of δ-catenin to postsynaptic sites through interaction with the Shank3 N-terminus.
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DOI：
10.1186/s13229-020-00385-8
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发表时间：
2020-10-28
期刊：
影响因子：
6.2
通讯作者：
Kreienkamp HJ
中科院分区：
文献类型：
--
作者：
Hassani Nia F;Woike D;Martens V;Klüssendorf M;Hönck HH;Harder S;Kreienkamp HJ
神经发育障碍如自闭症谱系障碍（ASD）可能是由编码参与突触形成和功能的蛋白质的基因的改变引起的。这包括支架蛋白如Shank 3和突触粘附蛋白如Neurexins或Neuroligins。一个重要的问题是个体风险基因的产物是否在功能上合作（例如Neurexin与Neuroligin亚型的相互作用）。这可能表明一个共同的发病途径。对于SHANK 3基因，在ASD病例中观察到杂合性功能丧失以及错义突变。几个错义突变影响Shank 3的N-末端部分，其包含高度保守的Shank/ProSAP N-末端（SPN）和锚蛋白重复（Ank）结构域。这些结构域的作用以及这些突变与Shank 3突触功能的相关性还很不清楚。我们使用纯化的突触蛋白馏分，以及各种生化和细胞生物学方法，以确定与Shank 3 N-末端在突触后位点的蛋白。我们在这里报告，δ-catenin，这是由CTNND 2，自闭症候选基因编码，直接与Ank结构域的Shank 3在突触后位点通过其Armadillo-repeat域相互作用。这种相互作用不受已知的δ-连环蛋白的翻译后修饰（即磷酸化或棕榈酰化）的影响。然而，Shank 3的SPN结构域中的ASD相关突变L 68 P显著增加了Shank 3与δ-连环蛋白的相互作用。通过对小鼠突触后组分的分析，我们表明缺乏含有SPN-Ank的Shank 3的大同种型导致突触后δ-连环蛋白的损失。此外，在原代培养的神经元中，含有N-末端结构域的Shank 3变体的表达显著增加了突触后位点共表达的δ-连环蛋白的存在。在模型生物如小鼠和原代培养的神经元中的工作可能无法忠实地再现人脑神经元中的情况。在原代培养的神经元中的工作也由于缺乏内源性δ-连环蛋白的特异性抗体而受到阻碍。我们的数据表明，Shank 3 N-末端和δ-catenin之间的相互作用是δ-catenin的突触后靶向所必需的。δ-连环蛋白未能正确靶向突触后位点可能导致自闭症谱系障碍的发病机制。
Neurodevelopmental disorders such as autism spectrum disorder (ASD) may be caused by alterations in genes encoding proteins that are involved in synapse formation and function. This includes scaffold proteins such as Shank3, and synaptic adhesion proteins such as Neurexins or Neuroligins. An important question is whether the products of individual risk genes cooperate functionally (exemplified in the interaction of Neurexin with Neuroligin isoforms). This might suggest a common pathway in pathogenesis. For the SHANK3 gene, heterozygous loss of function, as well as missense mutations have been observed in ASD cases. Several missense mutations affect the N-terminal part of Shank3 which contains the highly conserved Shank/ProSAP N-terminal (SPN) and Ankyrin repeat (Ank) domains. The role of these domains and the relevance of these mutations for synaptic function of Shank3 are widely unknown. We used purification from a synaptic protein fraction, as well as a variety of biochemical and cell biological approaches to identify proteins which associate with the Shank3 N-terminus at postsynaptic sites. We report here that δ-catenin, which is encoded by CTNND2, an autism candidate gene, directly interacts with the Ank domain of Shank3 at postsynaptic sites through its Armadillo-repeat domain. The interaction is not affected by well-known posttranslational modifications of δ-catenin, i.e. by phosphorylation or palmitoylation. However, an ASD-associated mutation in the SPN domain of Shank3, L68P, significantly increases the interaction of Shank3 with δ-catenin. By analysis of postsynaptic fractions from mice, we show that the lack of SPN-Ank containing, large isoforms of Shank3 results in the loss of postsynaptic δ-catenin. Further, expression of Shank3 variants containing the N-terminal domains in primary cultured neurons significantly increased the presence of coexpressed δ-catenin at postsynaptic sites. Work in model organisms such as mice, and in primary cultured neurons may not reproduce faithfully the situation in human brain neurons. Work in primary cultured neurons was also hampered by lack of a specific antibody for endogenous δ-catenin. Our data show that the interaction between Shank3 N-terminus and δ-catenin is required for the postsynaptic targeting of δ-catenin. Failure of proper targeting of δ-catenin to postsynaptic sites may contribute to the pathogenesis of autism spectrum disorder.
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影响因子：
30.8
作者：
Durand, Christelle M.;Betancur, Catalina;Bourgeron, Thomas
通讯作者：
Bourgeron, Thomas
影响因子：
3.7
作者：
Kelleher RJ 3rd;Geigenmüller U;Hovhannisyan H;Trautman E;Pinard R;Rathmell B;Carpenter R;Margulies D
通讯作者：
Margulies D
影响因子：
9.2
作者：
Israely, I;Costa, RM;Liu, X
通讯作者：
Liu, X
影响因子：
4.4
作者：
Medina, M;Marinescu, RC;Kosik, KS
通讯作者：
Kosik, KS
影响因子：
5.2
作者：
Boccuto, Luigi;Lauri, Maria;Schwartz, Charles E.
通讯作者：
Schwartz, Charles E.