Drosophila S2 cells secrete wingless on exosome-like vesicles but the wingless gradient forms independently of exosomes.
Drosophila S2 cells secrete wingless on exosome-like vesicles but the wingless gradient forms independently of exosomes.
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DOI：
10.1111/tra.12016
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发表时间：
2013-01
期刊：
影响因子：
--
通讯作者：
Vincent JP
中科院分区：
文献类型：
--
作者：
Beckett K;Monier S;Palmer L;Alexandre C;Green H;Bonneil E;Raposo G;Thibault P;Le Borgne R;Vincent JP
无翼基因在果蝇的翅盘中起着形态原的作用，它指定细胞命运，并控制离其表达位点数个细胞直径的生长。因此，尽管被酰化和膜相关，无翼细胞在细胞外空间扩散。最近的研究集中在确定Wingless在分泌途径中遵循的途径，并确定如何包装释放。我们发现，在由表达无翼果蝇S2细胞调节的培养基中，无翼细胞存在于外泌体样囊泡上，并且该部分激活信号转导。蛋白质组学分析表明，果蝇的无翅外泌体样结构含有许多与哺乳动物外泌体蛋白同源的蛋白质。此外，Evi，一种多通道跨膜蛋白，也存在于外泌体样囊泡中。利用这些外泌体标记和基于细胞的RNAi分析，我们发现小GTPase Rab11对外泌体的产生有重要贡献。这一发现使我们从体内Rab11敲除实验中得出结论，外泌体不太可能促进翅盘无翼分泌和梯度形成。与这一结论一致的是，细胞外标记的Evi从BAC中表达，不会从表达无翼细胞的想象盘释放。然而，Evi存在于幼虫的血淋巴中，这表明外泌体通常在体内产生。
Wingless acts as a morphogen in Drosophila wing discs, where it specifies cell fates and controls growth several cell diameters away from its site of expression. Thus, despite being acylated and membrane-associated, Wingless spreads in the extracellular space. Recent studies have focussed on identifying the route that Wingless follows in the secretory pathway and determining how it is packaged for release. We have found that, in medium conditioned by Wingless-expressing Drosophila S2 cells, Wingless is present on exosome-like vesicles and that this fraction activates signal transduction. Proteomic analysis shows that Wingless-containing exosome-like structures contain many Drosophila proteins that are homologous to mammalian exosome proteins. In addition, Evi, a multipass transmembrane protein, is also present on exosome-like vesicles. Using these exosome markers and a cell-based RNAi assay, we found that the small GTPase Rab11 contributes significantly to exosome production. This finding allows us to conclude from in vivo Rab11 knockdown experiments, that exosomes are unlikely to contribute to Wingless secretion and gradient formation in wing discs. Consistent with this conclusion, extracellularly tagged Evi expressed from a BAC is not released from imaginal disc Wingless-expressing cells. Nevertheless, Evi is present in larval haemolymph, suggesting that exosomes are normally produced in vivo.
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影响因子：
7.3
作者：
Baena-Lopez LA;Franch-Marro X;Vincent JP
通讯作者：
Vincent JP
影响因子：
11.4
作者：
Katanaev, Vladimir L.;Solis, Gonzalo P.;Hausmann, George;Buestorf, Silke;Katanayeva, Natalya;Schrock, Yvonne;Stuermer, Claudia A. O.;Basler, Konrad
通讯作者：
Basler, Konrad
影响因子：
2.6
作者：
GONZALEZ, F;SWALES, L;ARIAS, AM
通讯作者：
ARIAS, AM
DOI：
10.1083/jcb.97.2.329
发表时间：
1983-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Harding C;Heuser J;Stahl P
通讯作者：
Stahl P
影响因子：
64.5
作者：
Greco, V;Hannus, M;Eaton, S
通讯作者：
Eaton, S