Zhang Yan, Forinash Kara D, McGivern Jered, Fritz Brian, Dorey Karel, Sheets Michael D
University of Wisconsin-Madison, Department of Biomolecular Chemistry, 1300 University Avenue, Madison, WI 53706, USA.
Mol Cell Biol. 2009 Jul;29(13):3791-802. doi: 10.1128/MCB.01865-08. Epub 2009 Apr 13.
The xCR1 protein is a maternal determinant and cofactor for nodal signaling in vertebrate embryos. The xCR1 protein accumulates specifically in the animal cells of Xenopus embryos, but maternal xCR1 mRNA is distributed equally throughout all embryonic cells. Here, we show that vegetal cell-specific translational repression of xCR1 mRNA contributes to this spatially restricted accumulation of the xCR1 protein in Xenopus embryos. xCR1 mRNA was associated with polyribosomes in animal cells but not vegetal cells. A 351-nucleotide region of xCR1 mRNA's 3' untranslated region was sufficient to confer a spatially restricted pattern of translation to a luciferase reporter mRNA by repressing translation in vegetal cells. Repression depended upon the mRNA's 5' cap but not its 3' poly(A) tail. Furthermore, the region of xCR1 mRNA sufficient to confer vegetal cell-specific repression contained both Pumilio binding elements (PBEs) and binding sites for the CUG-BP1 protein. The PBEs and the CUG-BP1 sites were necessary but not sufficient for translation repression. Our studies of xCR1 mRNA document the first example of spatially regulated translation in controlling the asymmetric distribution of a maternal determinant in vertebrates.
xCR1蛋白是脊椎动物胚胎中节点信号传导的母体决定因素和辅助因子。xCR1蛋白特异性地积累在非洲爪蟾胚胎的动物细胞中,但母体xCR1 mRNA在所有胚胎细胞中均匀分布。在这里,我们表明xCR1 mRNA在植物细胞中的特异性翻译抑制导致了xCR1蛋白在非洲爪蟾胚胎中的这种空间受限积累。xCR1 mRNA在动物细胞中与多核糖体相关,但在植物细胞中不相关。xCR1 mRNA 3'非翻译区的一个351个核苷酸的区域足以通过抑制植物细胞中的翻译,赋予荧光素酶报告基因mRNA一种空间受限的翻译模式。抑制作用依赖于mRNA的5'帽而不是其3'聚腺苷酸尾。此外,足以赋予植物细胞特异性抑制作用的xCR1 mRNA区域同时包含Pumilio结合元件(PBEs)和CUG-BP1蛋白的结合位点。PBEs和CUG-BP1位点对于翻译抑制是必要的,但不是充分的。我们对xCR1 mRNA的研究记录了脊椎动物中空间调控翻译在控制母体决定因素不对称分布方面的首个例子。