STRUCTURE AND EXPRESSION OF EUCARYOTIC GENES

我们建议继续我们对结构和 线虫肌动蛋白基因的表达 优雅女装。我们现在将重点放在肌动蛋白信使RNA(Mrna)上。 加工和扩展工作，以包括加工其他 MRNAs。在研究肌动蛋白基因时，我们最近发现 四个基因中的三个编码经历了 MRNAs的5‘22个核苷酸发生不寻常的反应 来自单独的RNA分子。C。其他mRNAs。 优雅的人也会经历这种反式剪接反应。我们会研究 这一现象及其生物学意义。 我们将阐明肌动蛋白反式剪接的机制 线虫中的mRNAs。我们将对肌动蛋白的信使核糖核酸进行表征 前体、剪接前导RNA前体和分子 中间体。我们将使用体外剪接反应来鉴定 反应的成分，决定了单独的反式和 存在顺式元件(SnRNP和剪接体)，并且顺式剪接是 保持与反式剪接的分离，我们将识别其他mRNA 它们都经历了同样的反转录反应。我们会调查的 剪接前导是否赋予mRNA稳定性，增强 翻译，或本地化mRNA。我们将进行实验 以确定是否通过以下途径获得类似的剪接引导子 在其他真核生物中的反式剪接。

We propose to continue our studies on the structure and expression of the actin genes of the nematode Caenorhabditis elegans. We will now focus on actin messenger RNA (mRNA) processing and extend the work to include processing of other mRNAs. While studying the actin genes, we recently discovered that three of the four genes encode mRNAs that undergo an unusual reaction in which the 5' 22 nucleotides of the mRNAs are derived from a separate RNA molecule. Other mRNAs in C. elegans also undergo this trans-splicing reaction. We will study this phenomenon and its biological implications. We will elucidate the mechanism of trans-splicing of the actin mRNAs in C. elegans. We will characterize the actin mRNA precursors, the splice leader RNA precursor, and molecular intermediates. We will use in vitro splicing reactions to identify the components of the reaction, determine if separate trans-and cis- elements (snRNPs and spliceosomes) exist and cis-splicing is kept separate from trans-splicing we will identify other MRNAs that undergo the same transplicing reaction. We will investigate whether the splice leader confers mRNA stability, enhances translation, or localizes mRNAs. We will carry out experiments to determine whether similar splice leaders are acquired through trans-splicing in other eukaryotes.