NUCLEAR ROLE OF YEAST POLY(A)-BINDING PROTEIN

酵母多聚 (A) 结合蛋白的核作用

基本信息

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
项目类别：
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

项目摘要

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（改编自申请人的摘要）：拟议的研究旨在了解 Poly(A) 结合蛋白的多种功能与大多数真核生物 mRNA 的 Poly(A) 尾相关。有充足的有证据表明 Poly(A) 尾及其结合蛋白的存在对基因的表达具有深远的影响。这种RNA/蛋白质复合物刺激翻译起始并促进 mRNA 稳定细胞质。在细胞核中，mRNA 3' 端需要 Poly(A) 结合蛋白新生 mRNA 的形成和输出至细胞质。这个提议，使用酿酒酵母作为模型系统，解决了 Poly(A) 结合蛋白 (Pab1p) 的核功能。因素已发现与 Pab1p 一起调节聚腺苷酸化的程度。如果编码这些因子中最明确特征的基因 PBP1 是破坏后，poly(A) 尾部的长度显着减少在无细胞提取物中合成并抑制体内利用 HIS4 报告基因中的早期聚腺苷酸化位点。 PBP1 也被破坏抑制 PAB1 缺失的致死率，而不直接影响 mRNA 稳定性或翻译，这与表型形成对比先前分离出的pab1抑制器。两种杂交分析表明 Pbp1p 可能是 RNA 加工中招募的几个调节因素之一设备。研究Pbp1及相关因素在调控中的作用 Pab1p 在 3' 端处理中的功能 PI 建议：1) 分析 Pbp1p 和其他 Pbp 的生化活性，包括评估 Pbp1p 调节聚腺苷酸核酸酶或聚腺苷酸活性的能力聚合酶； 2) 鉴定其表达依赖于相关基因的mRNA 使用寡核苷酸探针阵列测定聚腺苷酸化的程度； 3）分析通过确定 PAB1 和 PBP1 的相互作用来确定 PAB1 和 PBP1 的遗传关系与编码切割和/或多腺苷酸化因子的其他基因表征生长缺陷的高复制抑制因子这些基因的过度表达。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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：
发表时间：
2004-07
期刊：
影响因子：
0
作者：
Parker, Kenneth C;Patterson, Dale;Williamson, Brian;Marchese, Jason;Graber, Armin;He, Feng;Jacobson, Allan;Juhasz, Peter;Martin, Stephen
通讯作者：
Martin, Stephen

Identification of factors regulating poly(A) tail synthesis and maturation.

鉴定调节 Poly(A) 尾合成和成熟的因素。