POST-TRANSCRIPTIONAL CONTROL OF THE HEART AND LUNGS

（改编自申请人摘要）从基因转录的各种RNA 与心血管和肺部疾病有关的是在 转录水平。 这些包括β-肾上腺素能受体，β 2- 肾上腺素能受体、血管内皮生长因子（VEGF）和 白细胞介素-3（IL-3）mRNA。 这些转录本含有富含腺苷/尿苷的 在它们的3'非翻译区（3'-UTR）中的元件（战神），其在以下中起作用： 调节它们的稳定性和翻译。 战神靶向mRNA， 衰变，通常通过poly（A）缩短依赖性衰变途径。 的mRNA 战神的去稳定化、poly（A）缩短和翻译功能是 由ARE结合蛋白（ARE-BPs）介导。 有趣的是， 控制这些转录物的丰度会导致疾病。 对这些mRNA的药理学操作需要了解这些基因的表达。 调节其基因表达的分子机制。 最伟大的 然而，在研究战神如何与ARE-BP相互作用以 调节基因表达。 然而，在研究如何 战神与ARE-BP相互作用以调节基因表达是能够 证明所确定的ARE-BP实际上是后 特定转录物的转录调控。 我们开发了一个 系统研究ARE介导的mRNA衰变途径。 插入 将肿瘤坏死因子α（TNF α）ARE插入酵母的3 'UTR MFA 2 mRNA引起嵌合mRNA的快速降解。 AUF 1的表达， 酿酒酵母中的ARE-BP特异性地影响 这种含有ARE的mRNA表明酵母系统重现了 在哺乳动物细胞中获得的结果。 这个实验的目的是 赠款提案是利用酵母的遗传和分子方面， 在酵母和哺乳动物细胞中研究ARE介导的mRNA衰变途径。 具体来说，我们建议：a）研究β- 肾上腺素能受体，IL-3和VEGF战神促进mRNA周转使用酵母 酿酒酵母作为模型系统; B）研究这些战神如何调节 其mRNA在哺乳动物细胞和无细胞系统中的稳定性;和c） 开发分析以表征β-肾上腺素能受体的作用， IL-3和VEGF战神翻译。 这些研究将帮助我们扩大我们的 了解战神和ARE-BP，以及这些相互作用如何控制 参与心血管和肺部疾病的mRNA表达，以及 如癌症和免疫系统紊乱。

(Adapted from applicant's abstract) Various RNAs transcribed from genes implicated in cardiovascular and pulmonary diseases are regulated at the post- transcriptional level. These include the beta-adrenergic receptor, the beta2- adrenergic receptor, the vascular endothelial growth factor (VEGF) and interleukin-3 (IL-3) mRNAs. These transcripts harbor Adenosine/Uridine-rich elements (AREs) in their 3' untranslated regions (3'-UTR) which play a role in regulating their stability and translation. AREs target mRNAs for rapid decay, usually via a poly (A) shortening-dependent decay pathway. The mRNA destabilizing, poly(A) shortening and translational functions of the AREs are mediated by ARE-binding proteins (ARE-BPs). Interestingly, inappropriate control of the abundance of these transcripts leads to disease. Pharmacological manipulation of these mRNAs requires an understanding of the molecular mechanisms regulating their gene expression. The greatest difficulty, however, in studying how the AREs interact with ARE-BPs to regulate gene expression. The greatest difficulty, however, in studying how the AREs interact with ARE-BPs to regulate gene expression is to be able to demonstrate that the identified ARE-BP is actually required for post- transcriptional regulation of a particular transcript. We have develop a system to investigate the ARE-mediated mRNA decay pathway in yeast. Insertion of the Tumor Necrosis Factor alpha(TNFalpha) ARE into the 3'UTR of the yeast MFA2 mRNA, causes rapid degradation of the chimeric mRNA. Expression of AUF1, an ARE-BP, in the yeast S.cerevisiae specifically affects the decay rate of this ARE-containing mRNA suggesting that the yeast system recapitulates the results obtained in mammalian cells. The goals of the experiments in this grant proposal are to utilize the genetic and molecular aspects of yeast to investigate the ARE-mediated mRNA decay pathway in yeast and mammalian cells. Specifically, we propose to: a) investigate the mechanism of how the beta- adrenergic receptors, IL-3 and VEGF AREs promote mRNA turnover using the yeast S.cerevisiae as a model system; b) investigate how these AREs regulate the stability of their mRNAs in mammalian cells and in cell-free system; and c) develop assays to characterize the effects of the beta-adrenergic receptors, IL-3 and VEGF AREs on translation. These studies will help us expand our knowledge on AREs and ARE-BPs, and how these interactions control the expression of mRNAs involved in cardiovascular and pulmonary diseases, as well as in cancers and immune disorders.