WDR35 is involved in subcellular localization of acetylated tubulin in 293T cells
WDR35 is involved in subcellular localization of acetylated tubulin in 293T cells
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DOI：
10.1016/j.bbrc.2021.01.092
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发表时间：
2021-02-18
影响因子：
3.1
通讯作者：
Furuno, Nobuaki
中科院分区：
文献类型：
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作者：
Sekiguchi, Takeshi;Ishii, Takashi;Furuno, Nobuaki
WDR35/IFT121是初级纤毛的纤束内转运蛋白，与mtorc1激活蛋白RagA相关。为了阐明WDR35和RagA在初级纤毛中的相互作用以及mTOR信号传导的功能，我们使用质谱法鉴定了WDR35相互作用蛋白。我们发现WDR35与CCT复合物蛋白相关，包括TCP1/CCT1，它们作为α -微管蛋白折叠的分子伴侣。免疫染色显示293T细胞乙酰化α -微管蛋白集中在原代纤毛附近。相比之下，乙酰化的微管蛋白分散在293T细胞构建的WDR35部分敲除细胞中。同样，在RagA敲除细胞中也观察到分散的乙酰化微管蛋白亚细胞定位。RagA存在于NIH3T3细胞的原毛中，并且RagA的GDP形式与WDR35有较强的结合，并负向调控原毛的形成。这些结果表明，WDR35通过与TCP1和/或RagA家族蛋白的相互作用参与初级纤毛乙酰化微管蛋白的亚细胞定位。(C) 2021爱思唯尔公司版权所有。
WDR35/IFT121 is an intraflagellar transport protein in primary cilia, which is associated with RagA, an mTORC1-activating protein. To elucidate the functions of the interaction between WDR35 and RagA in primary cilia, as well as mTOR signaling, we identified WDR35-interacting proteins using mass spectrometry. We found that WDR35 associates with CCT complex proteins including TCP1/CCT1, which act as molecular chaperones for alpha-tubulin folding. Immunostaining showed that acetylated alpha-tubulin was concentrated in the vicinity of primary cilia in 293T cells. In contrast, acetylated tubulin was dispersed in WDR35 partial knockout cells established from 293T cells. Similarly, scattered subcellular localization of acetylated tubulin was observed in RagA knockout cells. RagA was present in the primary cilia of NIH3T3 cells, and the GDP form of RagA exhibited strong binding to WDR35 and negative regulation of primary cilium formation. These results suggest that WDR35 is involved in the subcellular localization of acetylated tubulin in primary cilia via its interactions with TCP1 and/or RagA family proteins. (C) 2021 Elsevier Inc. All rights reserved.