Bauermeister Diana, Claußen Maike, Pieler Tomas
Department of Developmental Biochemistry, Göttingen Center for Molecular Biosciences (GZMB), University of Göttingen, D-37077 Göttingen, Germany.
Dev Biol. 2015 Sep 15;405(2):214-24. doi: 10.1016/j.ydbio.2015.07.005. Epub 2015 Jul 9.
The localization of certain mRNAs to the vegetal cortex of Xenopus oocytes is of crucial importance for germ cell development and early embryonic patterning. Vegetal RNA localization is mediated by cis-acting RNA localization elements (LE). Several proteins assemble on the RNA LE and direct transport to the vegetal cortex. Although a number of localization RNP components have been identified, their full composition is unknown. In an RNA affinity purification approach, using the dead end 1 (dnd1) RNA LE, we identified Xenopus Celf1 as a novel component of vegetal localization RNP complexes. Celf1 is part of an RNP complex together with known vegetal localization factors and shows specific interactions with LEs from several but not all vegetally localizing RNAs. Immunostaining experiments reveal co-localization of Celf1 with vegetally localizing RNA and with known localization factors. Inhibition of Celf1 protein binding by localization element mutagenesis as well as Celf1 overexpression interfere with vegetal RNA localization. These results argue for a role of Celf1 in vegetal RNA localization during Xenopus oogenesis.
某些mRNA在非洲爪蟾卵母细胞植物皮质的定位对于生殖细胞发育和早期胚胎模式形成至关重要。植物RNA定位由顺式作用RNA定位元件(LE)介导。几种蛋白质在RNA LE上组装并将其运输至植物皮质。尽管已鉴定出许多定位核糖核蛋白(RNP)成分,但其完整组成仍未知。在一种RNA亲和纯化方法中,我们利用死亡相关蛋白1(dnd1)RNA LE鉴定出非洲爪蟾Celf1是植物定位RNP复合物的一个新成分。Celf1与已知的植物定位因子一起构成RNP复合物的一部分,并与几种(但不是全部)植物定位RNA的LE显示出特异性相互作用。免疫染色实验揭示了Celf1与植物定位RNA以及已知定位因子的共定位。通过定位元件诱变抑制Celf1蛋白结合以及Celf1过表达均会干扰植物RNA定位。这些结果表明Celf1在非洲爪蟾卵子发生过程中的植物RNA定位中发挥作用。