TRIM9-dependent ubiquitination of DCC constrains kinase signaling, exocytosis, and axon branching.
TRIM9-dependent ubiquitination of DCC constrains kinase signaling, exocytosis, and axon branching.
复制标题
DCC的TRIM9依赖性泛素化限制了激酶信号传导，胞吐作用和轴突分支。
DOI：
10.1091/mbc.e16-08-0594
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发表时间：
2017-09-01
影响因子：
3.3
通讯作者：
Gupton SL
中科院分区：
文献类型：
--
作者：
Plooster M;Menon S;Winkle CC;Urbina FL;Monkiewicz C;Phend KD;Weinberg RJ;Gupton SL
在netrin的存在下，三联体基序蛋白9（TRIM 9）促进结肠直肠癌（DCC）中缺失的聚簇，但DCC的TRIM 9依赖性泛素化减少。泛素化的缺失促进DCC和FAK之间的相互作用和FAK活化。FAK激活是SNARE组装到胞吐囊泡融合的过程所必需的，胞吐囊泡融合为轴突分支提供膜材料。在结直肠癌（DCC）中缺失的细胞外netrin-1及其受体促进发育中的皮层神经元的轴突分支。Netrin依赖的形态发生之前是DCC的多聚化，FAK和Src家族激酶的激活，以及胞吐囊泡融合的增加，但这些事件是如何联系在一起的尚不清楚。在这里，我们证明了三重基序蛋白9（TRIM 9）依赖的DCC的泛素化阻断与FAK的相互作用和磷酸化。在netrin-1刺激后，TRIM 9促进DCC多聚化，但DCC的TRIM 9依赖性泛素化减少，这促进了与FAK的相互作用和随后的FAK活化。我们发现，抑制FAK活性阻断了体外胞吐频率的升高和体外和体内轴突分支的升高。虽然FAK抑制降低可溶性N-乙基马来酰亚胺附着蛋白受体（SNARE）介导的胞吐，组装的SNARE复合物和囊泡邻近质膜增加，表明FAK在从组装的SNARE复合物到囊泡融合的进展中发挥新的作用。
In the presence of netrin, tripartite motif protein 9 (TRIM9) promotes deleted in colorectal cancer (DCC) clustering, but TRIM9-dependent ubiquitination of DCC is reduced. Loss of ubiquitination promotes an interaction between DCC and FAK and FAK activation. FAK activation is required for the progression from SNARE assembly to exocytic vesicle fusion, which supplies membrane material for axon branching. Extracellular netrin-1 and its receptor deleted in colorectal cancer (DCC) promote axon branching in developing cortical neurons. Netrin-dependent morphogenesis is preceded by multimerization of DCC, activation of FAK and Src family kinases, and increases in exocytic vesicle fusion, yet how these occurrences are linked is unknown. Here we demonstrate that tripartite motif protein 9 (TRIM9)-dependent ubiquitination of DCC blocks the interaction with and phosphorylation of FAK. Upon netrin-1 stimulation TRIM9 promotes DCC multimerization, but TRIM9-dependent ubiquitination of DCC is reduced, which promotes an interaction with FAK and subsequent FAK activation. We found that inhibition of FAK activity blocks elevated frequencies of exocytosis in vitro and elevated axon branching in vitro and in vivo. Although FAK inhibition decreased soluble N-ethylmaleimide attachment protein receptor (SNARE)-mediated exocytosis, assembled SNARE complexes and vesicles adjacent to the plasma membrane increased, suggesting a novel role for FAK in the progression from assembled SNARE complexes to vesicle fusion in developing murine neurons.