PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure.
PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure.
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DOI：
10.1038/s41467-021-23116-w
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发表时间：
2021-05-24
影响因子：
16.6
通讯作者：
Kaeser PS
中科院分区：
文献类型：
--
作者：
Emperador-Melero J;Wong MY;Wang SSH;de Nola G;Nyitrai H;Kirchhausen T;Kaeser PS
突触前神经末梢的活动区定义了神经递质释放的位点。它的蛋白质机器可以通过液-液相分离来组织，这是一种形成无膜亚细胞区室的机制。在这里，我们表明，活性区蛋白Liprin-α3在转染的HEK 293 T细胞中快速可逆地发生相分离。凝液形成由Liprin-α3 PKC-磷酸化丝氨酸-760触发，RIM和Munc 13共同募集到膜附着的凝液中。磷酸特异性抗体在转染细胞和小鼠脑组织中建立Liprin-α3丝氨酸-760的磷酸化。在新产生的Liprin-α2/α3双敲除小鼠的原代海马神经元中，RIM和Munc 13的突触水平降低，可释放囊泡池减少。Liprin-α3的重新表达恢复了这些突触前缺陷，而突变Liprin-α3磷酸化位点以消除相位凝聚阻止了这种拯救。最后，这些神经元中的PKC激活急剧增加RIM、Munc 13和神经递质释放，这取决于磷酸化Liprin-α3的存在。我们的研究结果表明，PKC介导的Liprin-α3磷酸化触发其相分离，并调节活性区的结构和功能。液-液相分离可能是突触前神经末梢的组织化机制。在这里，作者表明PKC介导的Liprin-α3磷酸化触发细胞系的相分离，并调节原代海马神经元的活性区结构和功能。
The active zone of a presynaptic nerve terminal defines sites for neurotransmitter release. Its protein machinery may be organized through liquid–liquid phase separation, a mechanism for the formation of membrane-less subcellular compartments. Here, we show that the active zone protein Liprin-α3 rapidly and reversibly undergoes phase separation in transfected HEK293T cells. Condensate formation is triggered by Liprin-α3 PKC-phosphorylation at serine-760, and RIM and Munc13 are co-recruited into membrane-attached condensates. Phospho-specific antibodies establish phosphorylation of Liprin-α3 serine-760 in transfected cells and mouse brain tissue. In primary hippocampal neurons of newly generated Liprin-α2/α3 double knockout mice, synaptic levels of RIM and Munc13 are reduced and the pool of releasable vesicles is decreased. Re-expression of Liprin-α3 restored these presynaptic defects, while mutating the Liprin-α3 phosphorylation site to abolish phase condensation prevented this rescue. Finally, PKC activation in these neurons acutely increased RIM, Munc13 and neurotransmitter release, which depended on the presence of phosphorylatable Liprin-α3. Our findings indicate that PKC-mediated phosphorylation of Liprin-α3 triggers its phase separation and modulates active zone structure and function. Liquid–liquid phase separation may be a mechanism for organizing the presynaptic nerve terminal. Here, the authors show that PKC-mediated phosphorylation of Liprin-α3 triggers phase separation in cell lines and modulates active zone structure and function in primary hippocampal neurons.
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影响因子：
16.2
作者：
de Jong APH;Roggero CM;Ho MR;Wong MY;Brautigam CA;Rizo J;Kaeser PS
通讯作者：
Kaeser PS
影响因子：
5.7
作者：
Kaeser PS;Regehr WG
通讯作者：
Regehr WG
影响因子：
11.4
作者：
Ashery, U;Varoqueaux, F;Rettig, J
通讯作者：
Rettig, J
影响因子：
4.8
作者：
Ko, J;Na, M;Kim, E
通讯作者：
Kim, E
影响因子：
16.2
作者：
Held, Richard G.;Liu, Changliang;Kaeser, Pascal S.
通讯作者：
Kaeser, Pascal S.