Post-transcriptional regulation by the YBX3 RNA-binding protein in skeletal muscle

骨骼肌中 YBX3 RNA 结合蛋白的转录后调节

基本信息

批准号：
10439013
负责人：
Amy M. Cooke
金额：
$ 42.12万
依托单位：
HAVERFORD COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10439013
关键词：
Address Aging Amino Acid Transporter Amino Acids Binding Biochemical Biochemistry Bioinformatics Biological Biological Assay Biological Process Biology Cell Culture Techniques Cell physiology Cells Complex Data Data Set Defect Development Disease Embryonic Development Epilepsy Event Family Functional disorder Genes Genetic Materials Genetic Transcription Genomics Goals Health Homeostasis Human Impairment Individual Inflammation JAK1 gene Lead Learning Light Link Malignant Neoplasms Mammalian Cell Maps Mass Spectrum Analysis Mediating Memory Messenger RNA Methods Modeling Molecular Muscle Muscle Fibers Muscular Atrophy Nerve Degeneration Obesity Outcome Pathway interactions Post-Transcriptional Regulation Process Protein Biosynthesis Protein Family Proteins RNA RNA Binding RNA metabolism RNA-Binding Proteins Regulation Research Research Personnel Role Skeletal Muscle System Therapeutic Tissues Transcript Translations Work age related base biological adaptation to stress cell type clinically relevant experimental study human disease innovation interdisciplinary approach mRNA Stability member multiple omics novel therapeutic intervention protein complex skeletal muscle differentiation stem cells

项目摘要

PROJECT SUMMARY/ABSTRACT Post-transcriptional control permeates biology from proliferation to development. RNA-binding proteins (RBPs) dictate which messenger (m)RNAs are regulated, and how, where, and when that regulation occurs. Their roles in biology are incontrovertible, and emphasized by RBP dysfunctions that cause disease, including cancer, obesity and muscular atrophies. The long-term goal is to understand how RNA-protein complexes dictate and respond to complex biological events to realize how defects in these complexes result in disease. The objective of this proposal is to understand the diverse post-transcriptional regulation of the RBP YBX3, and connect its regulation to key biological processes and disease. The central hypothesis, which was formulated based on previous findings and preliminary data, is that YBX3 post-transcriptionally regulates mRNAs via multiple mechanisms, and this control is required to maintain amino acid transport in skeletal muscle. A multi-disciplinary approach that combines biochemistry, “omics”, bioinformatics and mammalian cell culture to dissect how YBX3 regulates via diverse mechanisms, and the role for its post-transcriptional control of amino acid transporters in skeletal muscles. The rationale for the proposed work is that once the diverse post-transcriptional control mechanisms are understood, this can be used as a paradigm for other clinically relevant RBPs, and to potentially develop therapeutic strategies based on this regulation. The objective of this project will be accomplished by three specific aims: 1) Define how the modular domains of YBX3 contribute to post-transcriptional regulation. The working hypothesis is that the modular domains of YBX3 help determine the diverse regulatory outcomes. The investigators will modify a well-established biochemical assay to assay the regulatory contribution of each domain. 2) Identify YBX3-dependent complexes formed on specific mRNAs. The working hypothesis is that different complexes form on mRNAs that YBX3 either activates or represses. RNA pull-down approaches will be used to identify transcript specific complexes using targeted and non-targeted methods. 3) Characterize the role of YBX3's post-transcriptional control of amino acid transport in skeletal muscle. The working hypothesis is that YBX3 stabilizes transporter mRNAs to maintain amino acid homeostasis in skeletal muscle that is critical for differentiation in this tissue. Amino acid mRNA stability, the intracellular levels of amino acids and differentiation will be assessed when YBX3 is depleted in skeletal muscle cells. This proposal is innovative because it 1) establishes how a single RBP controls mRNAs via multiple mechanisms, which can be used as a paradigm for other multi-regulatory RBPs, and 2) defines how post-transcriptional regulation impacts amino acid transport in skeletal muscle that could lead to new therapeutic strategies of a process that is impaired with aging. The proposed work is significant because it will 1) provide a model of how discrete domains regulate, 2) identify mRNA specific complexes required for regulation and 3) uncover how post-transcriptional control regulates amino acid transport and differentiation in skeletal muscle.

项目摘要/摘要转录后控制渗透到从增殖到发育的生物学。 RNA结合蛋白（RBP）决定调节哪些使者（M）RNA，以及该调节发生的方式，何时何地。他们的角色在生物学中是无可争议的，并且由RBP功能障碍强调，这些功能障碍引起癌症，包括癌症，肥胖和肌肉萎缩。长期目标是了解RNA蛋白复合物如何决定和对复杂的生物事件做出反应，以实现这些复合物中的缺陷如何导致疾病。目标该提议的内容是了解RBP YBX3的潜水员转录后调节调节关键生物过程和疾病。中央假设是基于以前的发现和初步数据是，转录后YBX3通过多个调节mRNA 机制，并且需要这种控制才能维持骨骼肌中的氨基酸转运。多学科结合了生物化学，“法律”，生物信息学和哺乳动物细胞培养的方法，以剖析YBX3 通过各种机制进行调节，以及其在氨基酸转运蛋白转录后的作用骨骼肌。拟议工作的理由是，一旦潜水员的转录后控制理解了机制，这可以用作其他临床相关的RBP的范例，并有可能根据该法规制定理论策略。该项目的目的将由三个具体目的：1）定义YBX3的模块化结构域如何促进转录后调节。工作假设是YBX3的模块化结构域有助于确定潜水员的调节结果。调查人员将修改建立良好的生化测定法，以主张每种测定领域。 2）确定在特定mRNA上形成的YBX3依赖性复合物。工作的假设是在mRNA上形成了不同的复合物，即YBX3激活或再现。 RNA下拉方法将是用于使用针对性和非靶向方法识别转录本特定的复合物。 3）表征角色 YBX3对骨骼肌中氨基酸转运的转录后控制。工作的假设是 YBX3稳定转运蛋白mRNA以维持骨骼肌中的氨基酸稳态，这对于该组织中的分化。氨基酸mRNA稳定性，氨基酸的细胞内水平和分化将YBX3耗尽在骨骼肌细胞中时，将评估。该提议具有创新性，因为它1）建立单个RBP如何通过多种机制控制mRNA，可以用作范式其他多调节RBP，以及2）定义转录后调节如何影响氨基酸的转运骨骼肌可能会导致新的治疗策略，从而受到衰老受损的过程。这提议的工作很重要，因为它将1）提供一个模型，说明离散域的调节方式，2）确定调节所需的mRNA特定复合物和3）揭示转录后控制如何调节骨骼肌中的氨基酸转运和分化。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The YBX3 RNA-binding protein posttranscriptionally controls SLC1A5 mRNA in proliferating and differentiating skeletal muscle cells.

DOI：
10.1016/j.jbc.2023.105602
发表时间：
2024-02
期刊：
JOURNAL OF BIOLOGICAL CHEMISTRY
影响因子：
4.8
作者：
Awad, Silina;Skipper, William;Vostrejs, William;Ozorowski, Kendall;Min, Kristen;Pfuhler, Liva;Mehta, Darshan;Cooke, Amy
通讯作者：
Cooke, Amy