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

繁殖所需的保守 RNA 结合蛋白的结构到表型分析

基本信息

批准号：
10798788
负责人：
Francesca Massi
金额：
$ 18.46万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-08-31
项目状态：
已结题

项目摘要

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.

在后生动物的发育过程中，体轴的极化是至关重要的，因为体轴不对称，细胞分裂启动细胞特化途径。一个保守的RNA结合蛋白家族其特征在于CCCH型串联锌指（TZF）结构域是轴极化所需的，细胞类型特化在C.优美的一系列对卵母细胞至关重要的事件- 向胚胎的转变由该家族的两个成员OMA-1/2启动。该过程开始于 OMA-1/2周转，并以细胞成分不对称分离达到高潮，胚胎的两极。OMA-1/2突变阻断卵母细胞成熟并阻止受精。我们已经观察到OMA-1在体外以相对低的特异性结合RNA，但调节特异性RNA，体内mRNA。我们还确定OMA-1 RNA结合是高度合作的，以前从未在CCCH型TZF家族的蛋白质中观察到。我们的目标是调查OMA- 1/2活性从结构到表型。我们将确定分子机制或RNA结合利用核磁共振光谱学、计算机模拟和定量生物化学的协同性。我们将还定义了RNA结合协同性如何定义OMA-1在蠕虫中的生物活性。我们将确定卵母细胞中哪些mRNAs受OMA-1/2的调控，以及这些mRNAs中哪些是对繁殖至关重要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。