Nucleation of the destruction complex on the centrosome accelerates degradation of β-catenin and regulates Wnt signal transmission.
Nucleation of the destruction complex on the centrosome accelerates degradation of β-catenin and regulates Wnt signal transmission.
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DOI：
10.1073/pnas.2204688119
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发表时间：
2022-09-06
影响因子：
11.1
通讯作者：
Wilson, Maxwell Z.
中科院分区：
文献类型：
--
作者：
Lach, Ryan S.;Qiu, Chongxu;Kajbaf, Erfan Zeyaei;Baxter, Naomi;Han, Dasol;Wang, Alex;Lock, Hannah;Chirikian, Orlando;Pruitt, Beth;Wilson, Maxwell Z.
液-液相分离（LLPS）控制着各种中尺度细胞过程。然而，我们对细胞如何利用LLPS来驱动细胞功能知之甚少。在这里，我们研究了破坏复合体（DC），这是一种控制Wnt信号的细胞器，其成分相分离。通过先进的显微镜，CRISPR，计算模型和光遗传学的结合，我们发现DC由中心体成核，并且这种成核驱动有效的信号转导。我们的工作不仅揭示了LLPS的生物学功能，而且强调了成核作为控制细胞内凝聚物功能的一般方法。最后，我们的研究结果表明，Wnt信号转导与细胞周期之间存在热力学耦合，这可能有助于深入了解Wnt驱动的癌症。Wnt信号转导由破坏复合体（DC）控制，DC是一种包含支架蛋白和调节β-连环蛋白稳定性的激酶的凝聚物。过表达的DC支架经过液-液相分离（LLPS），但内源性表达水平的DC中尺度组织及其在β-catenin加工中的作用此前尚不清楚。这里，我们发现DC LLPS是由中心体成核的。通过结合crispr工程定制荧光标签，有限元模拟和光遗传学工具，允许操纵DC浓度和多价，我们发现中心体成核通过将DC组分共定位到单个反应坩埚中来驱动β-catenin的加工。在中心体上富集GSK3β可控制β-catenin加工并阻止wnt驱动的胚胎干细胞向中胚层分化。我们的研究结果证明了核子在控制生物分子凝聚中的作用，并表明Wnt信号转导与细胞周期之间存在紧密的联系。
Liquid–liquid phase separation (LLPS) governs a variety of mesoscale cellular processes. However, less is known about how cells utilize LLPS to drive cellular function. Here, we examined the destruction complex (DC), an organelle which controls Wnt signaling and whose components phase separate. Through a combination of advanced microscopy, CRISPR, computational modeling, and optogenetics, we find that the DC is nucleated by the centrosome and that this nucleation drives efficient signal transduction. Our work not only uncovers a biological function for LLPS but also highlights nucleation as a general method for controlling the function of intracellular condensates. Finally, our findings suggest a thermodynamic coupling between Wnt signal transduction and the cell cycle which could lead to insights into Wnt-driven cancers. Wnt signal transduction is controlled by the destruction complex (DC), a condensate comprising scaffold proteins and kinases that regulate β-catenin stability. Overexpressed DC scaffolds undergo liquid–liquid phase separation (LLPS), but DC mesoscale organization at endogenous expression levels and its role in β-catenin processing were previously unknown. Here, we find that DC LLPS is nucleated by the centrosome. Through a combination of CRISPR-engineered custom fluorescent tags, finite element simulations, and optogenetic tools that allow for manipulation of DC concentration and multivalency, we find that centrosomal nucleation drives processing of β-catenin by colocalizing DC components to a single reaction crucible. Enriching GSK3β partitioning on the centrosome controls β-catenin processing and prevents Wnt-driven embryonic stem cell differentiation to mesoderm. Our findings demonstrate the role of nucleators in controlling biomolecular condensates and suggest tight integration between Wnt signal transduction and the cell cycle.
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影响因子：
56.9
作者：
Bashor, Caleb J.;Helman, Noah C.;Lim, Wendell A.
通讯作者：
Lim, Wendell A.
DOI：
10.1073/pnas.1400568111
发表时间：
2014-04-15
影响因子：
11.1
作者：
Bazzi, Hisham;Anderson, Kathryn V.
通讯作者：
Anderson, Kathryn V.
DOI：
10.1073/pnas.1509317112
发表时间：
2015-09-22
影响因子：
11.1
作者：
Berry, Joel;Weber, Stephanie C.;Brangwynne, Clifford P.
通讯作者：
Brangwynne, Clifford P.
影响因子：
3.3
作者：
Elmore, Zachary C.;Guillen, Rodrigo X.;Gould, Kathleen L.
通讯作者：
Gould, Kathleen L.
影响因子：
9.2
作者：
Cliffe, A;Hamada, F;Bienz, M
通讯作者：
Bienz, M