Institute for Genetics, Heinrich-Heine-University Düsseldorf, D-40225 Düsseldorf, Germany.
Dev Biol. 2010 Mar 15;339(2):268-79. doi: 10.1016/j.ydbio.2009.12.002. Epub 2009 Dec 11.
Understanding how epithelial organs form during morphogenesis is a major problem in developmental biology. In the present paper, we provide a detailed analysis of vang-1, the only homolog of the planar cell polarity protein Strabismus/Van Gogh in Caenorhabditis elegans. We demonstrate that during organogenesis of the intestine, (i) VANG-1 specifically interacts with PDZ 2 domain of DLG-1 (Discs large) and becomes phosphorylated by the kinase domain of the FGF-like receptor tyrosine kinase EGL-15; (ii) VANG-1 is predominantly restrained to the cell cortex but relocates to the apical junction; and (iii) in vang-1 embryos epithelial cells of the intestine are not correctly arranged along the anterior-posterior axis. To investigate what determines the disposition of the VANG-1 protein, either truncated protein forms were expressed in the intestine or RNAi was used to remove the functions of gene products previously shown to be involved in apical junction formation. Removal of the VANG-1 PDZ binding motif "-ESAV" and depletion of dlg-1 or let-413 gene functions interferes with the localization of VANG-1. In addition, egl-15 embryos show a premature relocation of VANG-1 to the apical junction, causing defects that resemble those observed in mutant vang-1 embryos and after intestine-specific overexpression of full-length vang-1. Finally, the localization of VANG-1 depends on DSH-2, a homolog of the planar cell polarity protein Dishevelled and depletion phenocopies vang-1 and egl-15 phenotypes in the embryonic intestine.
在形态发生过程中理解上皮器官的形成是发育生物学中的一个主要问题。在本文中,我们对 vang-1 进行了详细分析,vang-1 是秀丽隐杆线虫中平面细胞极性蛋白 Strabismus/Van Gogh 的唯一同源物。我们证明,在肠的器官发生过程中,(i)VANG-1 特异性地与 PDZ2 结构域的 DLG-1(Discs large)相互作用,并被 FGF 样受体酪氨酸激酶 EGL-15 的激酶结构域磷酸化;(ii)VANG-1 主要被限制在细胞膜皮质,但重新定位到顶端连接;(iii)在 vang-1 胚胎中,肠上皮细胞没有沿着前后轴正确排列。为了研究什么决定了 VANG-1 蛋白的分布,我们在肠中表达了截短的蛋白形式,或者使用 RNAi 去除以前被证明参与顶端连接形成的基因产物的功能。去除 VANG-1 的 PDZ 结合基序“-ESAV”和 dlg-1 或 let-413 基因功能的缺失会干扰 VANG-1 的定位。此外,egl-15 胚胎中 VANG-1 过早地重新定位到顶端连接,导致类似于突变 vang-1 胚胎和肠特异性全长 vang-1 过表达后观察到的缺陷。最后,VANG-1 的定位取决于 DSH-2,DSH-2 是平面细胞极性蛋白 Dishevelled 的同源物,其缺失表型类似于 vang-1 和 egl-15 在胚胎肠中的表型。
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