NUCLEAR ROLE OF YEAST POLY(A)-BINDING PROTEIN

酵母多聚 (A) 结合蛋白的核作用

• 批准号: 6636425
• 负责人: Allan S Jacobson
• 金额: $ 24.78万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6636425
• 关键词: RNA binding protein; Saccharomyces cerevisiae; enzyme activity; fungal genetics; fungal proteins; gene interaction; messenger RNA; microarray technology; monoclonal antibody; nucleic acid probes; phenotype; polyadenylate; suppressor mutations; yeast two hybrid system

DESCRIPTION (Adapted from applicant's abstract): The proposed research seeks to understand the diverse functions of the poly(A)-binding protein that is associated with the poly(A) tail of most eukaryotic mRNAs. There is ample evidence indicating that the presence of the poly(A) tail and its bound protein has profound consequences on the expression of genes. This RNA/protein complex stimulates initiation of translation and promotes mRNA stabilization in the cytoplasm. In the nucleus, poly(A)-binding protein is required for mRNA 3'-end formation and export of the nascent mRNA from to the cytoplasm. This proposal, using the yeast Saccharomyces cerevisiae as a model system, addresses the nuclear functions of poly(A)-binding protein (Pab1p). Factors have been identified that, in concert with Pab1p, regulate the extent of polyadenylation. If the gene encoding the most well-characterized of these factors, PBP1, is disrupted, there is a substantial reduction in the lengths of poly(A) tails synthesized in cell-free extracts and inhibits the in vivo utilization of an early polyadenylation site in a HIS4 reporter. Disruption of PBP1 also suppresses the lethality of a PAB1 deletion without directly effecting mRNA stability or translation, which stands in contrast to the phenotypes of previously isolated pab1 suppressors. Two-hybrid analyses suggest that Pbp1p may be one of several regulatory factors recruited to the RNA processing apparatus. To study the role of Pbp1 and related factors in the regulation of Pab1p's function in 3'-end processing the PI proposes to: 1) analyze the biochemical activities of Pbp1p and the other Pbps, including an assessment of Pbp1p's ability to regulate the activity of poly(A) nuclease or poly(A) polymerase; 2) identify mRNAs whose expression is dependent on the relative extent of polyadenylation using oligonucleotide probe arrays; and 3) analyze the genetic relationships of PAB1 and PBP1 by determining their interactions with other genes encoding cleavage and/or polyadenylation factors by characterizing high copy suppressors of the growth defects resulting from overexpression of these genes.

描述(改编自申请者的摘要)：拟议的研究旨在 了解Poly(A)结合蛋白的不同功能 与大多数真核mRNAs的Poly(A)尾巴相关。有足够的 有证据表明Poly(A)尾巴及其结合蛋白的存在 对基因的表达有深远的影响。这种RNA/蛋白质复合体 刺激翻译的启动和促进mRNA的稳定 细胞质。在细胞核中，mRNA3‘端需要Poly(A)结合蛋白 新生信使核糖核酸的形成和输出到细胞质。这项提议， 使用酵母酿酒酵母作为模型系统，解决了 聚(A)结合蛋白(Pab1p)的核功能。因素一直是 发现与Pab1p共同调节多聚腺苷酸化的程度。 如果编码这些因子中最具特征的基因PBP1是 破坏后，Poly(A)尾巴的长度显著缩短 在无细胞提取物中合成，并抑制体内对AN的利用 HIS4报告中的早期多聚腺苷酸化位点。PBP1的中断也 在不直接影响mRNA的情况下抑制PAB1缺失的致命性 稳定性或平移性，这与 先前分离的pab1抑制子。双杂交分析表明，Pbp1p 可能是RNA加工过程中的几个调节因素之一 仪器。研究Pbp1及其相关因素在血管紧张素转换酶调控中的作用 Pab1p在3‘端加工中的作用PI提出：1)分析 Pbp1p和其他多溴联苯的生化活性，包括对 Pbp1p调节聚(A)核酸酶或聚(A)的活性 聚合酶；2)确定其表达依赖于亲缘关系的mRNAs 使用寡核苷酸探针阵列的多聚腺苷酸化程度；以及3)分析 通过确定PAB1和PBP1的相互作用来研究它们的遗传关系 与其他编码切割和/或多腺苷酸化因子的基因通过 高复制抑制子的生长缺陷的特征 这些基因的过度表达。

项目成果

Poly(A)-binding proteins: multifunctional scaffolds for the post-transcriptional control of gene expression.

poly（a）结合蛋白：用于基因表达的转录后控制的多功能支架。

• DOI: 10.1186/gb-2003-4-7-223
• 发表时间: 2003
• 期刊: GENOME BIOLOGY
• 影响因子: 12.3
• 作者: Mangus, David A;Evans, Matthew C;Jacobson, Allan
• 通讯作者: Jacobson, Allan

Depth of proteome issues: a yeast isotope-coded affinity tag reagent study.

蛋白质组问题的深度：酵母同位素编码的亲和标签试剂研究。

• DOI: 10.1074/mcp.m300110-mcp200
• 发表时间: 2004
• 期刊: Molecular & cellular proteomics : MCP
• 作者: Parker,KennethC;Patterson,Dale;Williamson,Brian;Marchese,Jason;Graber,Armin;He,Feng;Jacobson,Allan;Juhasz,Peter;Martin,Stephen
• 通讯作者: Martin,Stephen

Replacement of the yeast TRP4 3' untranslated region by a hammerhead ribozyme results in a stable and efficiently exported mRNA that lacks a poly(A) tail.

用锤头核酶替换酵母 TRP4 3 非翻译区会产生稳定且有效输出的 mRNA，该 mRNA 缺乏 Poly(A) 尾。

• DOI: 10.1017/s1355838202021039
• 发表时间: 2002
• 期刊: RNA (New York, N.Y.)
• 作者: Düvel,Katrin;Valerius,Oliver;Mangus,DavidA;Jacobson,Allan;Braus,GerhardH
• 通讯作者: Braus,GerhardH