Vazquez-Pianzola Paula, Schaller Bogdan, Colombo Martino, Beuchle Dirk, Neuenschwander Samuel, Marcil Anne, Bruggmann Rémy, Suter Beat
a Institute of Cell Biology, University of Bern , Bern , Switzerland.
b Interfaculty Bioinformatics Unit and Swiss Institute of Bioinformatics, University of Bern , Bern , Switzerland.
RNA Biol. 2017 Jan 2;14(1):73-89. doi: 10.1080/15476286.2016.1251542. Epub 2016 Nov 1.
mRNA (mRNA) transport focuses the expression of encoded proteins to specific regions within cells providing them with the means to assume specific functions and even identities. BicD and the mRNA binding protein Egl interact with the microtubule motor dynein to localize mRNAs in Drosophila. Because relatively few mRNA cargos were known, we isolated and identified Egl::GFP associated mRNAs. The top candidates were validated by qPCR, in situ hybridization and genetically by showing that their localization requires BicD. In young embryos these Egl target mRNAs are preferentially localized apically, between the plasma membrane and the blastoderm nuclei, but also in the pole plasm at the posterior pole. Egl targets expressed in the ovary were mostly enriched in the oocyte and some were apically localized in follicle cells. The identification of a large group of novel mRNAs associated with BicD/Egl points to several novel developmental and physiological functions of this dynein dependent localization machinery. The verified dataset also allowed us to develop a tool that predicts conserved A'-form-like stem loops that serve as localization elements in 3'UTRs.
信使核糖核酸(mRNA)运输将编码蛋白的表达集中于细胞内的特定区域,使其具备承担特定功能甚至特性的方式。在果蝇中,BicD和mRNA结合蛋白Egl与微管动力蛋白动力蛋白相互作用,以使mRNA定位。由于已知的mRNA货物相对较少,我们分离并鉴定了与Egl::GFP相关的mRNA。通过定量聚合酶链反应(qPCR)、原位杂交以及通过证明它们的定位需要BicD进行遗传学验证,确定了最有可能的候选者。在年轻胚胎中,这些Egl靶标mRNA优先定位于顶端,位于质膜和胚盘细胞核之间,但也存在于后极的极质中。在卵巢中表达的Egl靶标大多富集于卵母细胞中,有些则定位于卵泡细胞的顶端。与BicD/Egl相关的一大组新mRNA的鉴定,指出了这种依赖动力蛋白的定位机制的几种新的发育和生理功能。经过验证的数据集还使我们能够开发一种工具,该工具可预测保守的A'型茎环,这些茎环在3'非翻译区(3'UTR)中作为定位元件。