Temporal Dynamics and Stoichiometry in Notch Signaling - from Notch Synaptic Complex Formation to NICD Nuclear Entry.
Temporal Dynamics and Stoichiometry in Notch Signaling - from Notch Synaptic Complex Formation to NICD Nuclear Entry.
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Notch 信号传导中的时间动力学和化学计量学 - 从 Notch 突触复合体形成到 NICD 核进入。
DOI：
10.1101/2023.09.27.559780
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Blacklow,StephenC
中科院分区：
文献类型：
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作者：
Tveriakhina,Lena;Scanavachi,Gustavo;Egan,EmilyD;Correia,RicardoBangoDaCunha;Martin,AlexandreP;Rogers,JuliaM;Yodh,JeremyS;Aster,JonC;Kirchhausen,Tom;Blacklow,StephenC
当Delta或Jagged与Notch结合刺激Notch细胞内结构域（NICD）的蛋白水解释放时，就会发生哺乳动物Notch信号传导，后者进入细胞核以调节靶基因表达。为了确定在天然条件下与Notch信号传导相关的事件的时间动态，我们在Notch和Delta的内源基因组基因座处荧光标记Notch和Delta，并在接收器（Notch）和发送器（Delta）细胞配对后将其可视化为细胞接触后时间的函数。在接触部位，Notch和Delta立即以1：1的化学计量在突触中积累，其在接触后15-20分钟消退。突触的形成先于Notch细胞外结构域进入发送细胞和NICD在接收细胞的细胞核中的积累，其在45分钟后接近最大值，并且被信号传导的化学和遗传抑制剂阻止。这些发现以前所未有的时空精度将Notch-Delta突触动力学与NICD产生直接联系起来。
Mammalian Notch signaling occurs when binding of Delta or Jagged to Notch stimulates proteolytic release of the Notch intracellular domain (NICD), which enters the nucleus to regulate target gene expression. To determine the temporal dynamics of events associated with Notch signaling under native conditions, we fluorescently tagged Notch and Delta at their endogenous genomic loci and visualized them upon pairing of receiver (Notch) and sender (Delta) cells as a function of time after cell contact. At contact sites, Notch and Delta immediately accumulated at 1:1 stoichiometry in synapses, which resolved by 15-20 min after contact. Synapse formation preceded entrance of the Notch extracellular domain into the sender cell and accumulation of NICD in the nucleus of the receiver cell, which approached a maximum after ∼45 min and was prevented by chemical and genetic inhibitors of signaling. These findings directly link Notch-Delta synapse dynamics to NICD production with unprecedented spatiotemporal precision.