FUNCTION OF 3 UTRS

3个UTRS的功能

• 批准号: 6385883
• 负责人: Marvin P. Wickens
• 金额: $ 31.78万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6385883
• 关键词: Xenopus oocyte; cell cycle; cell cycle proteins; embryo /fetus; fertilization; gene expression; genetic regulation; genetic translation; messenger RNA; nucleic acid sequence; polyadenylate; posttranscriptional RNA processing

Eukaryotic mRNAs can be controlled at many different steps. In the nucleus, transcription and mRNA processing are required to produce an mRNA that can be translated. In the cytoplasm, mature mRNAs can be regulated at the level of stability, transnational activity, and cellular location. The objective of the proposed research is to understand the biochemical mechanisms that control mature mRNAs in animal cells. We focus on the roles of poly (A) addition and removal in the transnational control of mRNAs during early development. These reactions are regulated by sequences that lie between the translation termination codon and the poly (A) tail--in the 3' untranslated region of the mRNA (3'UTR). Our ultimate goals are to understand, in molecular terms how poly (A) and elements in the 3'UTR control translation. We propose to study these reactions in frog oocytes and embryos. We focus on the period of oocyte maturation and use cycline and c-mos mRNAs, as models. We have shown that changes in poly (A) length can regulate translation during early development, and that sequences in the 3"UTR govern the extent and timing of transnational activation and repression. We have developed straightforward in vivo and in vitro assays for the effects of poly (A) and sequences in the 3"UTR. We propose to identify how poly (A) stimulates translation by combining in vivo and in vitro approaches. Using molecular genetics and biochemistry, we will identify the step(s) at which poly (A) exerts its effects and examine the role of the protein to which it binds. By constructing well-defined mutations in synthetic mRNAs, we will precisely identify elements in the 3"UTR that control transnational activity in vivo and determine how they function. We will test our hypothesis that changes in the length of the c-mos mRNA's poly (A) tail, governed by signals in its 3"UTR, are critical for control of the embryonic cell cycle. The work proposed will elucidate fundamental mechanisms of gene expression, and therefore has important practical applications. Regulated changes in poly (A) length occur in many, if not all, animal species, affect many mRNAs in the embryo, and continue throughout life. Our detailed investigations of the regulation of c-mos, a proto-oncogene normally expressed only in the germ line, bear on how cell division is controlled both in normal and abnormal growth.

真核 mRNA 可以通过许多不同的步骤进行控制。 在 细胞核、转录和 mRNA 加工是产生 可翻译的mRNA。 在细胞质中，成熟的 mRNA 可 在稳定性、跨国活动和细胞水平上进行监管 地点。 拟议研究的目的是了解生化 控制动物细胞中成熟 mRNA 的机制。 我们专注于 聚（A）添加和去除在跨国控制中的作用 早期发育期间的 mRNA。 这些反应受调节 位于翻译终止密码子和多聚密码子之间的序列 (A) 尾部——mRNA 的 3' 非翻译区 (3'UTR)。 我们的最终目标是从分子角度理解聚 (A) 和 3'UTR 中的元素控制翻译。 我们建议研究这些 青蛙卵母细胞和胚胎的反应。 我们关注的是卵母细胞的时期 成熟并使用 cycline 和 c-mos mRNA 作为模型。 我们已经证明，聚 (A) 长度的变化可以调节翻译 在早期发育过程中，3"UTR 中的序列控制着 跨国激活和镇压的程度和时间。 我们有 开发了直接的体内和体外测定法来检测 聚 (A) 和 3"UTR 中的序列。 我们建议通过结合来确定聚 (A) 如何刺激翻译 体内和体外方法。 利用分子遗传学和 生物化学中，我们将确定聚 (A) 发挥其作用的步骤 效应并检查其结合的蛋白质的作用。 经过 在合成 mRNA 中构建明确定义的突变，我们将精确地 确定 3"UTR 中控制跨国活动的要素 体内并确定它们如何发挥作用。 我们将检验我们的假设 c-mos mRNA 的聚 (A) 尾长度的变化，由以下因素控制 其 3"UTR 中的信号对于胚胎细胞的控制至关重要 循环。 拟议的工作将阐明基因的基本机制 表达，因此具有重要的实际应用价值。 多聚 (A) 长度的调节变化发生在许多（如果不是全部）动物身上 物种，影响胚胎中的许多 mRNA，并持续终生。 我们对原癌基因 c-mos 调控的详细研究 通常仅在生殖系中表达，与细胞分裂的方式有关 控制正常和异常生长。