Mapping the proximity interaction network of the Rho-family GTPases reveals signalling pathways and regulatory mechanisms
Mapping the proximity interaction network of the Rho-family GTPases reveals signalling pathways and regulatory mechanisms
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DOI：
10.1038/s41556-019-0438-7
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发表时间：
2019-12-23
影响因子：
21.3
通讯作者：
Cote, Jean-Francois
中科院分区：
文献类型：
--
作者：
Bagci, Halil;Sriskandarajah, Neera;Cote, Jean-Francois
鸟嘌呤核苷酸交换因子（RhoGEF）和GTP酶激活蛋白（RhoGAP）协调GTP酶的Rho家族的激活状态以结合效应物。在这里，我们利用邻近依赖性生物素化来系统地定义Rho家族邻近相互作用网络，从28个诱饵中产生9，939个高置信度的邻近相互作用。利用Rho GTP酶的核苷酸状态，我们揭示了与RhoGEF和RhoGAP相互作用的景观。我们系统地定义了Rho蛋白的效应子，以揭示经典和非典型Rho蛋白的候选者。我们使用光遗传学来证明KIAA 0355（这里称为GARRE）是RAC 1相互作用子。RHOG候选效应子的功能筛选鉴定PLEKHG 3为RHOG下游Rac介导的膜皱褶的启动子。我们发现，活性RHOA结合激酶SLK在果蝇和哺乳动物细胞，以促进Ezrin-Radixin-Moesin磷酸化。我们的邻近相互作用数据为解剖额外的Rho信号通路铺平了道路，这里描述的方法适用于Ras family.Using a BioID approach，Bagci et al.系统地分析了Rho家族GTdR相互作用组，并揭示了以前未被重视的与RhoGEFs和RhoGAP以及Rho蛋白效应子的相互作用。
Guanine nucleotide exchange factors (RhoGEFs) and GTPase-activating proteins (RhoGAPs) coordinate the activation state of the Rho family of GTPases for binding to effectors. Here, we exploited proximity-dependent biotinylation to systematically define the Rho family proximity interaction network from 28 baits to produce 9,939 high-confidence proximity interactions in two cell lines. Exploiting the nucleotide states of Rho GTPases, we revealed the landscape of interactions with RhoGEFs and RhoGAPs. We systematically defined effectors of Rho proteins to reveal candidates for classical and atypical Rho proteins. We used optogenetics to demonstrate that KIAA0355 (termed GARRE here) is a RAC1 interactor. A functional screen of RHOG candidate effectors identified PLEKHG3 as a promoter of Rac-mediated membrane ruffling downstream of RHOG. We identified that active RHOA binds the kinase SLK in Drosophila and mammalian cells to promote Ezrin-Radixin-Moesin phosphorylation. Our proximity interactions data pave the way for dissecting additional Rho signalling pathways, and the approaches described here are applicable to the Ras family.Using a BioID approach, Bagci et al. systematically analyse the Rho-family GTPase interactome and reveal previously unappreciated interactions with RhoGEFs and RhoGAPs and effectors for Rho proteins.