COORDINATION OF PROCESSING AND TRANSPORT OF MRNAS

MRNAS 处理和运输的协调

• 批准号: 2599811
• 负责人: Pamela A Silver
• 金额: $ 26.99万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2599811
• 关键词: arginine; chemical binding; enzyme activity; enzyme inhibitors; heterogeneous nuclear ribonucleoprotein; immunoprecipitation; intracellular transport; messenger RNA; methylation; methyltransferase; peptide library; polymerase chain reaction; posttranscriptional RNA processing; tissue /cell culture

The goal of the proposed research is to understand how processing and movement of messenger RNAs out of the nucleus is coordinated. Following their synthesis, mRNAs are subjected to extensive processing including capping, splicing, polyadenylation and packaging into protein-RNA particles for export out of the nucleus. An analysis of macromolecular trafficking across the nuclear envelope in the yeast Saccharomyces cerevisiae has revealed several key factors that play a role in mRNA processing and export out of the nucleus. These include: the NPL3 and HRP1 genes, which encode hnRNP proteins; HMT1, which encodes a novel protein methyltransferase and the CBP genes, which encode the major RNA cap binding proteins. The proposed experiments build on these observations and focus on three interrelated processes -binding of proteins to the 5' pre-mRNA cap, polyadenylation and export of mRNA through the nuclear pore and one enzyme - the novel arginine methyltransferase. The Specific Aims are: 1) to determine if there are additional arginine methyltransferases in yeast and to assess the reversibility of methylation on arginine; 2) to assess how mRNA polyadenylation is affected by protein methylation; 3) to define the relationship between polyadenylation factors and mRNA movement out of the nucleus; 4) to delineate the relationship between the RNA Cap Binding Complex (CBC) and mRNA export form the nucleus; and 5) to design novel inhibitors specific for arginine methyltransferases. Many viruses have been shown to exploit the endogenous nuclear import and export machinery in order to propagate. The finding that host proteins implicated in RNA export are, for example, methylated at arginine suggested that similar modifications might occur on viral proteins crucial for export of viral RNAs and this has been shown to be the case. An understanding of the role of methylation in cellular processes could lead to the discovery of new uses for methyltransferase inhibitors or inducers in, for example, treating viral infection. It is also of interest to note that both types of arginine-methylated proteins, hnRNPs and myelin basic protein, are prominent in autoimmune diseases; systemic lupus erythmatosis and multiple sclerosis, respectively. It may be that modified arginine has distinctive recognition properties that lead to initiation or exacerbation of autoimmune diseases. An understanding of the basic recognition properties of methylated arginine may be useful in finding out why such proteins are targets in autoimmune disease.

拟议研究的目的是了解处理方式和 Messenger RNA从细胞核中移动的移动是协调的。下列的 它们的合成，mRNA经过广泛的处理，包括 封盖，剪接，聚腺苷酸化和包装到蛋白质-RNA中 从细胞核中输出的颗粒。 大分子的分析 贩运酵母糖疗中的核包膜 酿酒酵母揭示了在mRNA中起作用的几个关键因素 从细胞核中加工和导出。 其中包括：NPL3和 HRP1基因，编码HNRNP蛋白； HMT1，编码小说 蛋白甲基转移酶和CBP基因，该基因编码主要RNA 盖结合蛋白。 拟议的实验以这些为基础 观察并关注三个相互关联的过程 - 结合 5'前MRNA帽的蛋白质，mRNA的聚腺苷酸化和导出 通过核孔和一种酶 - 新型精氨酸 甲基转移酶。 具体目的是：1）确定是否存在 酵母中的其他精氨酸甲基转移酶，并评估 精氨酸甲基化的可逆性； 2）评估mRNA的方式 聚腺苷酸受蛋白甲基化的影响； 3）定义 多腺苷酸化因子与mRNA移动之间的关系 核； 4）描述RNA帽之间的关系 结合复合物（CBC）和mRNA导出形成核； 5）设计 针对精氨酸甲基转移酶的新型抑制剂。 许多病毒已被证明可以利用内源性核进口 和导出机械以传播。 那个主人的发现 与RNA导出有关的蛋白质是例如在 精氨酸表明病毒可能会发生类似的修饰 蛋白质对于出口病毒RNA至关重要，这已被证明是 案件。 理解甲基化在细胞中的作用 过程可能导致发现甲基转移酶的新用途 例如，抑制剂或诱导剂治疗病毒感染。 它 同样值得注意的是，两种类型的精氨酸 - 甲基化 蛋白质，HNRNP和髓磷脂碱性蛋白质在自身免疫中突出 疾病；全身性狼疮红斑病和多发性硬化症， 分别。可能修改的精氨酸具有独特的 导致启动或加剧的识别属性 自身免疫性疾病。 对基本识别的理解 甲基化精氨酸的特性可能有助于找出为什么这样 蛋白质是自身免疫性疾病的靶标。