Structure to phenotype analysis of a conserved RNA binding protein required for reproduction

在后生动物的发育过程中，体轴的极化是至关重要的，因为它是不对称的 细胞分裂启动细胞特化途径。一个保守的RNA结合蛋白家族 以CCCH型串联锌指(TZF)结构域为特征的轴极化和 线虫早期胚胎的细胞类型规范。对卵母细胞至关重要的一系列事件- 到胚胎的转变是由该家族的两个成员OMA-1/2启动的。这个过程始于 OMA-1/2周转，并以细胞成分对相反方向的不对称分离而达到顶峰 胚胎的两极。OMA-1/2突变阻碍卵母细胞成熟并阻止受精。我们 已经观察到OMA-1在体外以相对较低的特异性结合RNA，但调节特定的 体内的mRNAs。我们还确定OMA-1 RNA结合是高度合作的，这是一个特征 以前从未在CCCH型TZF家族的蛋白质中发现过。我们的目标是调查OMA- 1/2活性从结构到表型。我们将确定RNA结合的分子机制 使用核磁共振光谱、计算机模拟和定量生物化学的协作性。我们会 还定义了RNA结合的协作性如何定义蠕虫中的OMA-1生物学活性。我们会 确定卵母细胞中哪些mRNAs受OMA-1/2调控，哪些mRNAs是 对繁殖最为关键。这项研究将导致对分子如何 OMA-1的特性决定了其在活体动物生殖生物学中的功能。

During metazoan development, polarization of the body axes is of critical importance, as asymmetric cell division initiates cell specialization pathways. A family of conserved RNA-binding proteins characterized by CCCH-type tandem zinc finger (TZF) domains is required for axis polarization and cell type specification in the early embryo of C. elegans. A cascade of events essential to the oocyte- to-embryo transition is initiated by two members of this family, OMA-1/2. The process initiates with OMA-1/2 turnover and culminates with asymmetric segregation of cellular components to opposing poles of the embryo. Mutation of OMA-1/2 blocks oocyte maturation and prevents fertilization. We have observed that OMA-1 binds RNA with relatively low specificity in vitro, but regulates specific mRNAs in vivo. We have also determined that OMA-1 RNA binding is highly cooperative, a feature never before observed in a protein from the CCCH-type TZF family. Our goal is to investigate OMA- 1/2 activity from structure to phenotype. We will determine the molecular mechanism or RNA-binding cooperativity using NMR spectroscopy, computer simulations, and quantitative biochemistry. We will also define how RNA-binding cooperativity defines OMA-1 biological activity in worms. We will determine which mRNAs are regulated by OMA-1/2 in oocytes and of these mRNAs which are the most critical to reproduction. This research will lead to a new understanding of how the molecular properties of OMA-1 determine its function in reproductive biology in living animals.