Endosome positioning coordinates spatially selective GPCR signaling.
Endosome positioning coordinates spatially selective GPCR signaling.
复制标题
内体定位协调空间选择性 GPCR 信号传导。
DOI：
10.1038/s41589-023-01390-7
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发表时间：
2024
影响因子：
14.8
通讯作者：
Tsvetanova,NikoletaG
中科院分区：
文献类型：
--
作者：
Willette,BlairKA;Zhang,Jin-Fan;Zhang,Jin;Tsvetanova,NikoletaG
G蛋白偶联受体（GPCR）可以启动独特的功能反应，这取决于亚细胞激活位点。努力揭示区室化GPCR信号传导的机制基础集中在这种调节的生物化学方面。在这里，我们评估的生物物理定位的受体含有内体作为一种替代的显着机制。我们设计了一种策略，以快速和选择性地重新分配含有受体的内体在完整的细胞中的“命令”，而不扰乱他们的生化组成。接下来，我们提出了两个互补的光学读数，使强大的测量批量和基因特异性GPCR/环AMP（cAMP）依赖的转录信号与单细胞分辨率。有了这些，我们确定天然内体定位的破坏抑制了内体依赖性转录反应的启动。最后，我们证明了一个突出的机制作用PDE介导的cAMP水解和本地蛋白激酶A活性在这个过程中。因此，我们的研究阐明了一种新的机制，通过确定内体定位作为空间选择性受体信号传导的主要介质来调节GPCR功能。
G-protein-coupled receptors (GPCRs) can initiate unique functional responses depending on the subcellular site of activation. Efforts to uncover the mechanistic basis of compartmentalized GPCR signaling have concentrated on the biochemical aspect of this regulation. Here we assess the biophysical positioning of receptor-containing endosomes as an alternative salient mechanism. We devise a strategy to rapidly and selectively redistribute receptor-containing endosomes ‘on command’ in intact cells without perturbing their biochemical composition. Next, we present two complementary optical readouts that enable robust measurements of bulk- and gene-specific GPCR/cyclic AMP (cAMP)-dependent transcriptional signaling with single-cell resolution. With these, we establish that disruption of native endosome positioning inhibits the initiation of the endosome-dependent transcriptional responses. Finally, we demonstrate a prominent mechanistic role of PDE-mediated cAMP hydrolysis and local protein kinase A activity in this process. Our study, therefore, illuminates a new mechanism regulating GPCR function by identifying endosome positioning as the principal mediator of spatially selective receptor signaling.