TRANSLATIONAL CONTROL OF GENE EXPRESSION BY MRNA

mRNA 对基因表达的翻译控制

• 批准号: 3283544
• 负责人: ROSEMARY JAGUS
• 金额: $ 11.33万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-12-01 至 1990-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3283544
• 关键词: Nematoda; adenosine triphosphate; antibody; binding proteins; calcium; cell free system; chemical binding; chemical structure function; crosslink; density gradient ultracentrifugation; developmental genetics; early embryonic stage; fertilization; gel electrophoresis; gene expression; genetic manipulation; genetic transcription; genetic translation; germ cells; histones; hydrolysis; immunochemistry; messenger RNA; nonmammalian vertebrate embryology; nuclease; nucleic acid probes; protein biosynthesis; protein kinase; protein structure; radiotracer; ribonucleoproteins; ribosomes; saltwater environment; sea urchins; synthetic nucleic acid

During oogenesis and maturation of the sea urchin egg, large stores of mRNA are accumulated for subsequent use during early embryogenesis. Utilization of stored maternal mRNA occurs at a very low rate prior to fertilization. After fertilization, dramatic changes in mRNA availability and recruitment occur, allowing rapid increases in the synthetic rate of many proteins during early cleavage. Changes have been documented in both overall rates of maternal mRNA utilization and in the selection of particular mRNA species at different developmental stages. Our previous studies have allowed the development of cell-free translation systems from eggs and embryos that reproduce the overall differences observed in the intact system. This investigation proposes to utilize these cell-free systems, in conjunction with other approaches, to identify the cellular components involved in the utilization of maternal mRNA. Using the cell-free systems an inhibitor of eIF-4F activity has been discovered in the unfertilized egg, that decreases gradually in activity over the first 2-3 hr after fertilization. The identity and nature of the eIF-4F inhibitory activity will be characterized. The mechanism by which this activity inhibits the initiation sequence will also be investigated. The manner in which the inhibitor is inactivated after fertilization will be explored. To complement these studies, a systematic search for developmentally regulated mRNA-interactive proteins will be undertaken, using a protein blotting and in vitro binding technique, with capped 32p-labelled mRNA's synthesized in vitro.

在海胆卵的发生和成熟过程中，大的 储存的信使核糖核酸在早期积累起来以供后续使用 胚胎发生。对存储的母体mRNA的利用发生在 受精率非常低。受精后，戏剧性的 MRNA的可获得性和募集发生变化，从而允许 许多蛋白质的合成速率在早期迅速增加 乳沟。在这两个总体比率中记录了变化 母体mRNA的利用及其在特异体选择中的作用 不同发育阶段的信使核糖核酸。我们以前的 研究使无细胞翻译的发展成为可能 来自卵子和胚胎的系统复制了整个 在完整的系统中观察到的差异。 这项调查建议利用这些无细胞系统，在 与其他方法相结合，以识别细胞 母体基因利用中涉及的成分。vbl.使用 无细胞系统是eIF-4F活性的抑制剂 在未受精卵中发现的，在 受精后2-3小时内的活动。身份和身份 将对EIF-4F抑制活性的性质进行表征。 这一活动抑制启动的机制 序列也将被调查。以一种方式 受精后的抑制物失活将被探索。至 作为这些研究的补充，系统地寻找 发育调节的mRNA相互作用蛋白将是 采用蛋白质印迹和体外结合 技术，用32P标记的信使核糖核酸体外合成。