Novel Regulatory Mechanisms of Drosophila Pumilio

果蝇 Pumilio 的新型调控机制

基本信息

批准号：
10312121
负责人：
Aaron Charles Goldstrohm
金额：
$ 32.12万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-20 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10312121
关键词：
Address Affect Ataxia Binding Biological Biological Assay Biological Process C-terminal Cancer Biology Cell Proliferation Cells Complex Data Defect Development Drosophila genus Embryo Embryonic Development Epilepsy Family Fertility Funding Gene Expression Gene Expression Regulation Genes Genetic Transcription Goals Health Human Human Biology Infertility Knowledge Malignant Neoplasms Mammals Maternal Messenger RNA Measures Mediating Memory Messenger RNA Modeling Molecular Motor N-terminal Nerve Degeneration Nervous System Physiology Nervous system structure Neurons Orthologous Gene Phenotype Poly(A)-Binding Proteins Post-Transcriptional Regulation Process Proteins RNA RNA Binding RNA Decay RNA Degradation RNA Recognition Motif RNA-Binding Proteins Regulation Repression Repressor Proteins Research Response Elements Role Specificity Structure System Testing Therapeutic Time Translational Repression Translations Work base egg genetic corepressor improved in vivo insight morphogens mutant nano nervous system disorder novel protein expression recruit stem cell proliferation stem cells

项目摘要

PROJECT SUMMARY The long term goal of our research is to determine how gene expression is regulated at the post- transcriptional level. Control of messenger RNA (mRNA) translation and degradation underlies important biological processes including development, fertility and neurological functions. The proposed work focuses on the archetypal mRNA regulator, Drosophila Pumilio (Pum), which belongs to a family of RNA-binding proteins that are conserved throughout eukarya. Pum binds an extensive group of mRNAs including those that encode key developmental morphogens. Upon binding to an mRNA, Pum represses expression of the encoded protein. We find that Pum accelerates decay of the target mRNA and the proposed research seeks to discover the mechanism of Pum-mediated mRNA degradation. We developed novel assays to measure Pum activity in Drosophila cells and discovered multiple unique Repression Domains that potently repress target mRNAs. In preliminary work, we identified key co-repressors that are necessary for Pum repression. In the first aim, we measure the impact of the Repression Domains on protein expression and mRNA degradation and interrogate the functional roles of two classes of mRNA decay factors in repression by each Pum Repression Domain. We interrogate the physical interactions of the co-repressors with each Pum Repression Domain and measure their recruitment to Pum-regulated target mRNAs. In the second aim, we investigate the role of each Pum Repression Domain in embryonic development and proper spatial and temporal control of gene expression during early development. This research will reveal novel mechanisms of Pum repression. The resulting discoveries are expected to broadly enhance our understanding of post-transcriptional control and specifically improve knowledge of gene regulation in development, fertility, stem cell proliferation, and the nervous system.

项目摘要我们研究的长期目标是确定如何在后期调节基因表达转录级别。 Messenger RNA（mRNA）翻译和降解的控制重要生物学过程，包括发育，生育和神经功能。拟议的工作重点是属于RNA结合蛋白家族的原型mRNA调节剂，果蝇Pumilio（PUM）在整个Eukarya中保守的。 PUM结合了一组广泛的mRNA，包括编码的mRNA 关键的发育形态剂。与mRNA结合后，PUM抑制编码的表达蛋白质。我们发现，PUM会加速目标mRNA的衰变，并提出的研究试图发现 PUM介导的mRNA降解的机制。我们开发了新颖的测定法以测量果蝇细胞并发现了多个有效抑制目标mRNA的独特抑制域。在初步工作，我们确定了pum抑制所必需的关键共抑制剂。在第一个目标中，我们测量抑制域对蛋白质表达和mRNA降解的影响每个PUM抑制域的抑制作用中两个类别的mRNA衰减因子的功能作用。我们询问共抑制剂与每个PUM抑制域的物理相互作用并测量他们招募了PUM调节的目标mRNA。在第二个目标中，我们研究了每个脓液的作用胚胎发育中的抑制域以及基因表达的适当空间和时间控制在早期发展期间。这项研究将揭示PUM抑制的新型机制。结果期望发现会广泛增强我们对转录后控制的理解，具体来说提高对发育，生育能力，干细胞增殖和神经系统的基因调节知识。

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Preparation of cooperative RNA recognition complexes for crystallographic structural studies.

用于晶体结构研究的协作 RNA 识别复合物的制备。

DOI：
10.1016/bs.mie.2019.04.001
发表时间：
2019
期刊：
Methods in enzymology
影响因子：
0
作者：
Qiu,Chen;Goldstrohm,AaronC;TanakaHall,TraciM
通讯作者：
TanakaHall,TraciM

A conserved domain of Drosophila RNA-binding protein Pumilio interacts with multiple CCR4-NOT deadenylase complex subunits to repress target mRNAs.

DOI：
10.1016/j.jbc.2022.102270
发表时间：
2022-09
期刊：
JOURNAL OF BIOLOGICAL CHEMISTRY
影响因子：
4.8
作者：
Haugen, Rebecca J.;Arvola, Rene M.;Connacher, Robert P.;Roden, Richard T.;Goldstrohm, Aaron C.
通讯作者：
Goldstrohm, Aaron C.

Human Pumilio proteins directly bind the CCR4-NOT deadenylase complex to regulate the transcriptome.

DOI：
10.1261/rna.078436.120
发表时间：
2021-04
期刊：
RNA (New York, N.Y.)
影响因子：
0
作者：
Enwerem III;Elrod ND;Chang CT;Lin A;Ji P;Bohn JA;Levdansky Y;Wagner EJ;Valkov E;Goldstrohm AC
通讯作者：
Goldstrohm AC

The RNA binding domain of Pumilio antagonizes poly-adenosine binding protein and accelerates deadenylation.