Evc2 is a positive modulator of Hedgehog signalling that interacts with Evc at the cilia membrane and is also found in the nucleus.
Evc2 is a positive modulator of Hedgehog signalling that interacts with Evc at the cilia membrane and is also found in the nucleus.
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DOI：
10.1186/1741-7007-9-14
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发表时间：
2011-02-28
期刊：
影响因子：
5.4
通讯作者：
Goodship JA
中科院分区：
文献类型：
--
作者：
Blair HJ;Tompson S;Liu YN;Campbell J;MacArthur K;Ponting CP;Ruiz-Perez VL;Goodship JA
Evc对软骨生长板中的印度刺猬（Hh）信号传导至关重要。编码Evc2的基因在不同的方向上与Evc非常接近，这两个人类基因的突变导致软骨发育不良Ellis-van Creveld综合征。生物信息学分析表明，Evc和Evc2基因在后生动物进化早期通过重复事件产生，随后在节肢动物和线虫中丢失。在这里，我们证明Evc2对于响应Smo激动剂嘌呤胺的Hh通路激活至关重要。以Evc为诱饵进行酵母双杂交筛选，发现Evc2是Evc的结合伙伴，并通过免疫沉淀法证实了相互作用。我们开发了抗Evc2抗体，并发现Evc2和Evc在基础体和初级纤毛上共定位。在转染的细胞中，当Evc和Evc2构建体同时转染时，可以观察到基体和纤毛的定位，而单独转染时则没有。我们发现Evc和Evc2是纤毛跨膜蛋白，其c端是细胞内和Evc2，但不是Evc，具有细胞外部分。此外，Evc2细胞中Evc在基底不存在。利用细胞质和核蛋白的Western blots，我们也证实全长Evc2位于细胞核中，而不是Evc。我们首次证明Evc2是Hh信号通路的正调节因子，并且它位于初级纤毛的基底。我们发现Evc和Evc2在基底和纤毛膜上的存在是相互依赖的。此外，细胞核中存在Evc2，但不存在Evc，这表明Evc2在纤毛和细胞核之间运动。
Evc is essential for Indian Hedgehog (Hh) signalling in the cartilage growth plate. The gene encoding Evc2 is in close proximity in divergent orientation to Evc and mutations in both human genes lead to the chondrodysplasia Ellis-van Creveld syndrome. Bioinformatic analysis reveals that the Evc and Evc2 genes arose through a duplication event early in metazoan evolution and were subsequently lost in arthropods and nematodes. Here we demonstrate that Evc2 is essential for Hh pathway activation in response to the Smo agonist purmorphamine. A yeast two-hybrid screen using Evc as bait identified Evc2 as an Evc binding partner and we confirmed the interaction by immunoprecipitation. We developed anti-Evc2 antibodies and show that Evc2 and Evc co-localize at the basal body and also on primary cilia. In transfected cells, basal body and cilia localization is observed when Evc and Evc2 constructs are co-transfected but not when either construct is transfected individually. We show that Evc and Evc2 are cilia transmembrane proteins, the C-terminus for both being intracellular and Evc2, but not Evc, having an extracellular portion. Furthermore, Evc is absent at the basal body in Evc2 null cells. Using Western blots of cytoplasmic and nuclear protein, we also demonstrate that full length Evc2 but not Evc, is located in the nucleus. We demonstrate for the first time that Evc2 is a positive regulator of the Hh signalling pathway and that it is located at the basal body of primary cilia. We show that the presence of Evc and Evc2 at the basal body and cilia membrane is co-dependent. In addition, Evc2, but not Evc, is present in the cell nucleus suggesting movement of Evc2 between the cilium and nucleus.
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影响因子：
5.3
作者：
Tompson, Stuart W. J.;Ruiz-Perez, Victor L.;Goodship, Judith A.
通讯作者：
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影响因子：
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