Cytoplasmic retention of a nucleocytoplasmic protein TBC1D3 by microtubule network is required for enhanced EGFR signaling.
Cytoplasmic retention of a nucleocytoplasmic protein TBC1D3 by microtubule network is required for enhanced EGFR signaling.
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微管网络对核细胞质蛋白 TBC1D3 的细胞质保留是增强 EGFR 信号转导所必需的
DOI：
10.1371/journal.pone.0094134
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Shen C
中科院分区：
文献类型：
--
作者：
He Z;Tian T;Guo D;Wu H;Chen Y;Zhang Y;Wan Q;Zhao H;Wang C;Shen H;Zhao L;Bu X;Wan M;Shen C
类人癌基因TBC 1D 3增强表皮生长因子受体（EGFR）信号传导并诱导细胞转化。然而，关于其时空调控和肿瘤发生的机制知之甚少。在目前的研究中，我们确定了微管亚基β-微管蛋白作为一个潜在的相互作用伙伴TBC 1D 3使用亲和纯化结合质谱分析。通过免疫共沉淀证实了TBC 1D 3与β-微管蛋白之间的相互作用。使用相同的方法，我们还发现TBC 1D 3与内源性α-微管蛋白（微管的另一个亚基）共沉淀。与这些结果一致，微管共沉淀试验表明TBC 1D 3与微管网络相关。TBC 1D 3的β-微管蛋白相互作用位点定位于TBC结构域C-末端附近的氨基酸286 - 353。在这些氨基酸内的缺失突变显示出消除TBC 1D 3与β-微管蛋白的相互作用。有趣的是，缺失突变导致TBC 1D 3从细胞质丝状和点状结构中完全丢失，TBC 1D 3反而出现在细胞核中。与此一致，野生型TBC 1D 3在用微管解聚剂诺考达唑处理的细胞中表现出相同的核质分布，表明微管网络与TBC 1D 3缔合并将其保留在细胞质中。我们进一步发现，β-微管蛋白相互作用的缺陷导致TBC 1D 3不能抑制c-Cbl募集和EGFR泛素化，最终导致EGFR降解和信号转导的失调。总之，这些研究表明了一种新的模型，微管网络通过微管蛋白二聚体/寡聚体与核胞质蛋白TBC 1D 3的相互作用来调节EGFR的稳定性和信号传导。
The hominoid oncogene TBC1D3 enhances epidermal growth factor receptor (EGFR) signaling and induces cell transformation. However, little is known regarding its spatio-temporal regulation and mechanism of tumorigenesis. In the current study, we identified the microtubule subunit β-tubulin as a potential interaction partner for TBC1D3 using affinity purification combined with mass spectrometry analysis. The interaction between TBC1D3 and β-tubulin was confirmed by co-immunoprecipitation. Using the same method, we also revealed that TBC1D3 co-precipitated with endogenous α-tubulin, another subunit of the microtubule. In agreement with these results, microtubule cosedimentation assays showed that TBC1D3 associated with the microtubule network. The β-tubulin-interacting site of TBC1D3 was mapped to amino acids 286∼353 near the C-terminus of the TBC domain. Deletion mutation within these amino acids was shown to abolish the interaction of TBC1D3 with β-tubulin. Interestingly, the deletion mutation caused a complete loss of TBC1D3 from the cytoplasmic filamentous and punctate structures, and TBC1D3 instead appeared in the nucleus. Consistent with this, wild-type TBC1D3 exhibited the same nucleocytoplasmic distribution in cells treated with the microtubule depolymerizing agent nocodazole, suggesting that the microtubule network associates with and retains TBC1D3 in the cytoplasm. We further found that deficiency in β-tubulin-interacting resulted in TBC1D3's inability to inhibit c-Cbl recruitment and EGFR ubiquitination, ultimately leading to dysregulation of EGFR degradation and signaling. Taken together, these studies indicate a novel model by which the microtubule network regulates EGFR stability and signaling through tubulin dimer/oligomer interaction with the nucleocytoplasmic protein TBC1D3.
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影响因子：
4.8
作者：
Carbonaro, Marisa;Escuin, Daniel;Giannakakou, Paraskevi
通讯作者：
Giannakakou, Paraskevi
影响因子：
3.7
作者：
Kong C;Samovski D;Srikanth P;Wainszelbaum MJ;Charron AJ;Liu J;Lange JJ;Chen PI;Pan ZQ;Su X;Stahl PD
通讯作者：
Stahl PD
DOI：
10.1111/j.1600-0854.2011.01305.x
发表时间：
2012-02
期刊：
Traffic (Copenhagen, Denmark)
影响因子：
--
作者：
Eden ER;Huang F;Sorkin A;Futter CE
通讯作者：
Futter CE
影响因子：
5.7
作者：
Hatanaka, Hisashi;Tsukui, Mamiko;Mano, Hiroyuki
通讯作者：
Mano, Hiroyuki
影响因子：
4.8
作者：
Bowers, K;Piper, SC;Luzio, JP
通讯作者：
Luzio, JP