PROTEIN ARGININE METHYLATION AND RNA MATURATION

蛋白质精氨酸甲基化和 RNA 成熟

• 批准号: 6151225
• 负责人: MICHAEL F HENRY
• 金额: $ 21.98万
• 依托单位: UNIV OF MED/DENT NJ-SCH OSTEOPATHIC MED
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6151225
• 关键词: RNA biosynthesis; Saccharomyces cerevisiae; arginine; biological signal transduction; chemical binding; fungal genetics; fungal proteins; heterogeneous nuclear ribonucleoprotein; high performance liquid chromatography; intracellular transport; matrix assisted laser desorption ionization; methylation; methyltransferase; posttranscriptional RNA processing; protein localization; protein sequence

The overall goal of the proposed research is to understand the physiological role of protein arginine methylation and its impact on the maturation of mRNA in the yeast Saccharomyces cerevisiae. Given the prevalence of this post-translational modification among RNA metabolism enzymes, it is likely to play an important role in the process. In exploring this hypothesis further, the first aim will be to analyze the impact of arginine methylation on Hrp 1p, an essential yeast protein that is a member of a class of proteins termed heterogeneous nuclear ribonucleoproteins or hnRNPs. To provide a foundation for this analysis, the dimethylarginine residues within Hrp 1p will first be mapped. Next, the effect of this modification on three Hrp 1p activities will be assayed; l) mRNA 3'-end processing, 2) RNA-binding, and 3) nucleocytoplasmic shuttling. The second aim examines whether protein arginine methylation represents a signaling mechanism by determining the reversibility of the process. If so, the plan is to identify and characterize the corresponding demethylase. The third aim utilizes a combination of biochemical and molecular biological approaches to identify and characterize proteins that physically interact or functionally overlap with the protein arginine methyltransferase enzyme (Hmt1p). As part of this aim, cellular proteins that receive this modification will be cataloged. The objective is to learn where these proteins are localized in the cell, and whether they share common sequences and cellular activities. Since the structures of arginine methyltransferases have been well conserved from yeast to human cells, the genetically amenable yeast Saccharomyces cerevisiae represents an ideal model organism for these studies. Recent data implicates some human arginine methylated proteins in molecular diseases (i.e. FMR1: Fragile X Mental Retardation Syndrome). Additionally, the presence of methylated arginines among viral RNA maturation enzymes implies a role for this modification in viral mRNA processing (i.e. herpes simplex ICP27). Furthermore, human arginine methylated hnRNPs participate RNA metabolism which impacts additional cellular processes. A better understanding of arginine methylation should not only enhance the understanding of cellular RNA maturation, but also provide insights into viral mechanisms for RNA maturation during cell infection.

这项研究的总体目标是了解蛋白质精氨酸甲基化在酿酒酵母中的生理作用及其对mRNA成熟的影响。鉴于这种翻译后修饰在RNA代谢酶中的普遍存在，它很可能在这一过程中发挥重要作用。在进一步探索这一假说时，第一个目标将是分析精氨酸甲基化对HRP 1p的影响，HRP 1p是一种基本的酵母蛋白，是一种被称为异质性核糖核蛋白或hnRNPs的蛋白质的成员。为了为这一分析提供基础，首先将绘制HRP 1p中的二甲基精氨酸残基。接下来，我们将检测这种修饰对三种HRP 1p活性的影响：L)mRNA3‘端加工，2)RNA结合，3)核质穿梭。第二个目的是通过确定该过程的可逆性来检验蛋白质精氨酸甲基化是否代表一种信号机制。如果是这样的话，我们的计划是鉴定和鉴定相应的脱甲基酶。第三个目的是利用生化和分子生物学相结合的方法来识别和表征与蛋白质精氨酸甲基转移酶(Hmt1p)物理上相互作用或功能上重叠的蛋白质。作为这一目标的一部分，接受这种修饰的细胞蛋白质将被编目。目的是了解这些蛋白质在细胞中的定位，以及它们是否具有共同的序列和细胞活动。由于精氨酸甲基转移酶的结构从酵母到人类细胞都被很好地保存了下来，因此遗传适应性酵母酿酒酵母是进行这类研究的理想模式生物。最近的数据表明，一些人类精氨酸甲基化蛋白与分子疾病(即FMR1：脆性X智力低下综合征)有关。此外，病毒RNA成熟酶中甲基化精氨酸的存在暗示了这种修饰在病毒mRNA处理中的作用(即单纯疱疹病毒ICP27)。此外，人类精氨酸甲基化的hnRNPs参与RNA代谢，从而影响额外的细胞过程。更好地理解精氨酸甲基化，不仅可以提高对细胞RNA成熟的理解，而且还可以为深入了解细胞感染过程中RNA成熟的病毒机制提供帮助。