The Talpid3 gene (KIAA0586) encodes a centrosomal protein that is essential for primary cilia formation
The Talpid3 gene (KIAA0586) encodes a centrosomal protein that is essential for primary cilia formation
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DOI：
10.1242/dev.028464
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发表时间：
2009-02-15
期刊：
影响因子：
4.6
通讯作者：
Tickle, Cheryll
中科院分区：
文献类型：
--
作者：
Yin, Yili;Bangs, Fiona;Tickle, Cheryll
鸡TALPID(3)突变体在KIAA0568中有突变，具有多指和依赖于刺猬(HH)信号的其他胚胎区域(如神经管)的缺陷。在小鼠和人类综合征中也可以看到类似的表型，编码中心体或鞭毛内运输蛋白的基因发生突变。这种突变会导致初级纤毛的缺陷，而初级纤毛是HH信号发生的地方。在这里，我们证明了Talpid(3)突变胚胎的细胞缺乏初级纤毛，并且初级纤毛可以通过编码Talpid3的构建物来挽救。塔尔比德突变胚胎还发育出多囊肾，这与普遍的纤毛形成失败相一致。突变型神经管的超微结构研究表明，基底体成熟，但未能与顶端细胞膜对接，方向错误，几乎完全缺乏纤毛轴丝。我们还在突变型细胞中发现了肌动蛋白组织的显著变化，这可能解释了基底定位错误的原因。KIAA0586在人中心体蛋白质组中被鉴定出来，我们用鸡Talpid3的抗体在野生型鸡细胞的中心体中检测到Talpid3，而在突变细胞中没有检测到Talpid3。海葵线虫Talpid3同源基因的克隆和生物信息学分析表明，Talpid3蛋白中有一个高度保守的区域，包括一个预测的卷曲结构域。我们表明，这个区域是挽救初级纤毛形成和神经管模式所必需的，并且足以进行中心体定位。因此，Talpid3是影响纤毛发生和HH信号的越来越多的中心体蛋白之一。
The chicken talpid(3) mutant, with polydactyly and defects in other embryonic regions that depend on hedgehog (Hh) signalling ( e. g. the neural tube), has a mutation in KIAA0568. Similar phenotypes are seen in mice and in human syndromes with mutations in genes that encode centrosomal or intraflagella transport proteins. Such mutations lead to defects in primary cilia, sites where Hh signalling occurs. Here, we show that cells of talpid(3) mutant embryos lack primary cilia and that primary cilia can be rescued with constructs encoding Talpid3. talpid(3) mutant embryos also develop polycystic kidneys, consistent with widespread failure of ciliogenesis. Ultrastructural studies of talpid(3) mutant neural tube show that basal bodies mature but fail to dock with the apical cell membrane, are misorientated and almost completely lack ciliary axonemes. We also detected marked changes in actin organisation in talpid(3) mutant cells, which may explain misorientation of basal bodies. KIAA0586 was identified in the human centrosomal proteome and, using an antibody against chicken Talpid3, we detected Talpid3 in the centrosome of wild-type chicken cells but not in mutant cells. Cloning and bioinformatic analysis of the Talpid3 homolog from the sea anemone Nematostella vectensis identified a highly conserved region in the Talpid3 protein, including a predicted coiled-coil domain. We show that this region is required to rescue primary cilia formation and neural tube patterning in talpid(3) mutant embryos, and is sufficient for centrosomal localisation. Thus, Talpid3 is one of a growing number of centrosomal proteins that affect both ciliogenesis and Hh signalling.