Complex Involved In mRNA Decay in Yeast

参与酵母 mRNA 衰变的复合物

• 批准号: 6941767
• 负责人: STUART W PELTZ
• 金额: $ 30.81万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6941767
• 关键词: SDS polyacrylamide gel electrophoresis; Saccharomyces cerevisiae; adenosine diphosphate; binding sites; fungal genetics; genetic transcription; immunoprecipitation; messenger RNA; microarray technology; nucleic acid metabolism; nucleic acid sequence; phosphorylation; posttranscriptional RNA processing; posttranslational modifications; protein protein interaction; stimulus /response; stress proteins; translation factor

DESCRIPTION (provided by applicant): The decay of mRNA is central to the post-transcriptional regulation of gene expression. The expression of many clinically relevant genes, including cytokines, proto-oncogenes and growth factors, is regulated at the level of mRNA turnover through AU-rich elements (AREs) in their 3' untranslated regions. Therefore, studies of this process provide valuable insight into the deregulation of cellular mechanisms in some cancers and immune disorders. This proposal aims to use the yeast, Saccharomyces cerevisiae, as a model system to study the phenomenon of ARE-mediated mRNA decay. The enzymes and pathways of mRNA turnover are well characterized in this organism. Regulation of AREmediated mRNA decay in yeast occurs in response to at least two cellular stimuli and involves three identified factors; Publp, Cthlp and Cth2p. The goals of this proposal are to determine how (i) the ARE-binding complex regulates mRNA decay rates (ii) the ARE-binding complex is modulated in response to cellular stimuli and (iii) the complex of factors that assembles on the ARE is specified its sequence context. The first part of the proposal concentrates on characterization of the three ARE-binding proteins and the interactions among them and between them and the ARE. One important aspect focuses on dissecting the interaction between the Cth proteins and Pablp. In addition, experiments in this Aim will determine how these factors are post-translationally modified in response to cellular conditions. Aim II of the proposal is to isolate ARE-binding complexes formed in vivo using both RNA tags and protein tags. Together the results of this part of the study will identify novel components of the ARE-binding complex and give us insight into how these complexes are altered in response to cellular conditions. The final Aim of the proposal is to identify novel AU-rich elements in yeast through microarray analysis of mRNA decay rates in strains lacking the known ARE-binding factors and by analysis of candidate ARE-containing transcripts discovered by searching the Transterm database. The goal is to amass a set of AU-rich elements large enough to facilitate computer analysis of the sequences to identify commonalities between them that might specify the proteins they interact with and their regulation.

描述（由申请人提供）：mRNA的衰减对于基因表达的转录后调节至关重要。许多临床相关基因的表达，包括细胞因子，原始基因和生长因子，在通过3'未翻译区域中通过Au富元素（ARE）的mRNA更新水平调节。因此，对这一过程的研究为某些癌症和免疫疾病中细胞机制的放松管制提供了宝贵的见解。该建议旨在使用酵母酿酒酵母作为模型系统，以研究介导的mRNA衰变现象。在这种生物体中，mRNA更新的酶和途径很好地表征了。对酵母中产生的mRNA衰变的调节是针对至少两个细胞刺激而发生的，涉及三个鉴定因子。 Publp，CTHLP和CTH2P。该提案的目标是确定（i）结合复合物如何调节mRNA衰减率（ii）响应细胞刺激和（iii）指定其序列上下文中组成的因子的复合物。提案的第一部分集中于三种结合蛋白的表征以及它们之间以及它们之间的相互作用。一个重要方面的重点是剖析CTH蛋白与PABLP之间的相互作用。此外，在此目标中的实验将决定这些因素在响应细胞条件的响应后翻译后如何修饰。该提案的目的II是使用RNA标签和蛋白质标签隔离在体内形成的结合复合物。总之，这一研究的结果将确定结合结合复合物的新成分，并让我们深入了解这些复合物如何响应细胞条件。该提案的最终目的是通过微阵列分析缺乏已知的是结合因子的菌株中的mRNA衰变率来鉴定酵母中新型的AU富含元素，并且通过搜索ternterm数据库发现的候选者的分析是候选者的分析。目的是积聚一组足够大的元素元素，以促进对序列的计算机分析，以识别它们之间的共同点，这些元素可以指定它们与之相互作用的蛋白质及其调节。