Molecular Genetics of Maternal RNA Regulation in C. elegans

母体rna的调控对早期发育至关重要。金布尔博士建议采用遗传和分子相结合的方法来分析秀丽隐杆线虫的母体RNA控制。在早期胚胎中，fem3和glp-1基因表达调控性别决定和诱导细胞相互作用的母体rna。初步证据表明，这些rna的3'非翻译区在翻译水平上调节它们的活性。Kimble博士提出通过三种方式来解决母体rna控制fem3和glp-1的机制。首先，她将询问母体的fam -3和glp-1 rna是否受到翻译控制。为此，她将研究它们的内源性rna和蛋白质的分布，并评估它们与多聚体的关联。其次，她将确定调控所需的序列要素。为此，她将携带野生型和突变型序列的报告rna从fem-3或glp-1注入种系，并检测其在卵母细胞和早期胚胎中的翻译能力。第三，她将开始分析调控母体rna的因素。为此，她将依靠凝胶移位法和mog-1突变体分析。了解秀丽隐杆线虫如何调节母体rna，将有助于深入了解这种对早期发育的基本控制，也将允许在早期胚胎中操纵特定蛋白质的表达。

The regulation of maternal RNAs is critical to early development. Dr. Kimble proposes to take a combined genetic and molecular approach to analyze maternal RNA controls in the nematode C. elegans. The fem-3 and glp-1 genes express maternal RNAs that regulate sex determination and inductive cell interactions in early embryos. Preliminary evidence suggests that the 3' untranslated regions of these RNAs regulate their activity at the level of translation. Dr. Kimble proposes to address the mechanisms by which the fem-3 and glp-1 maternal RNAs are controlled in three ways. First, she will ask whether the fem-3 and glp-1 maternal RNAs are translationally controlled. To this end, she will examine the distribution of their endogenous RNAs and proteins and assess their association with polysomes. Second, she will identify sequence elements necessary for regulation. To this end, she will inject reporter RNAs carrying wild-type and mutant sequences from fem-3 or glp-1 into the germ line and assay their translational capacity in oocytes and early embryos. Third, she will begin to analyze factors that regulate maternal RNAs. To this end, she will rely on gel shift assays and analysis of mog-1 mutants. An understanding of how maternal RNAs are regulated in C. elegans will provide insight into this fundamental control of early development and also will allow expression of specific proteins to be manipulated in early embryos.