Posttranscriptional regulation of the Wnt pathway by HuR and microRNAs

HuR 和 microRNA 对 Wnt 通路的转录后调控

基本信息

批准号：
7389000
负责人：
KYLE D MANSFIELD
金额：
$ 4.96万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2009-03-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Posttranscriptional regulation (PTR) has come into vogue since the discovery of microRNAs and their ability to regulate developmental processes. However, RNA-binding proteins (RBPs) are well-established regulators of mRNAs and also play major roles in modulating PTR. This proposal will examine the cooperation and/or competition between microRNAs and the Hu RBPs in regulating mRNAs encoding members of the Wnt/B-catenin Pathway. The Wnt pathway plays important roles in cell-cell communication during development and in tumors. Recently, we have identified 61 Wnt pathway members whose mRNA associate with HuR in Jurkat T cells. Given that the Wnt pathway has been heavily studied in neuronal systems, this proposal will examine the regulation of these mRNAs by Hu proteins and predicted microRNAs during neuronal differentiation of P19 embryonic carcinoma cells treated with retinoic acid. I hypothesize that Hu proteins collaborate with microRNAs during neuronal differentiation to regulate the production of Wnt pathway components, and that this results in coordination of expression per the posttranscriptional RNA operon (PTRO) model derived previously in the Keene laboratory. The specific aims of the project are: 1. Identify the mRNA targets associated with HuR during neuronal development. We will use ribonucleoprotein immunoprecipitation followed by microarray analysis (RIP-Chip) to identify subsets of messages associated with HuR and investigate their dynamics during P19 neuronal development. 2. Investigate the interaction between HuR and miRNAs in neurons. RIP-Chip analysis, along with real time PCR and Northern analysis will be used to identify the subset of miRNAs associated with the HuR mRNP in neurons. 3. Investigate HuR's posttranscriptional role during neuronal development. This aim will study the impact of HuR on the expression and regulation of the Wnt pathway during neuronal development. By overexpressing and/or knocking down HuR in P19 cells and looking at the effects on Wnt family member RNA and protein expression and transcriptional activity we can begin to understand the regulatory role of HuR. Relevance: This project will investigate the posttranscriptional regulation of the Wnt pathway during neuronal development. By better understanding this oft-overlooked form of regulation, we will identify potential targets for therapeutic intervention in diseases involving the Wnt pathway.

描述（由申请人提供）：自从发现microRNA及其调节发育过程的能力以来，转录后调节（PTR）已进入时尚。然而，RNA结合蛋白（RBP）是mRNA的完善调节剂，并且在调节PTR中也起着重要作用。该提案将研究microRNA和HU RBP之间在调节编码WNT/B-catenin途径成员的mRNA方面的合作和/或竞争。 WNT途径在发育和肿瘤过程中在细胞 - 细胞通信中起重要作用。最近，我们确定了61个Wnt途径成员，其mRNA与Jurkat T细胞中的HUR相关。鉴于Wnt途径在神经元系统中进行了大量研究，因此该建议将检查HU蛋白对这些mRNA的调节，并在p19胚胎癌细胞的神经元分化过程中预测用维黄酮酸治疗的mrot元。我假设HU蛋白在神经元分化过程中与microRNAS合作以调节Wnt途径成分的产生，并且这会导致根据先前在Keene实验室中得出的转录后RNA操纵子（PTRO）模型来协调表达。该项目的具体目的是：1。确定在神经元发育过程中与HUR相关的mRNA靶标。我们将使用核糖核蛋白免疫沉淀，然后进行微阵列分析（RIP-CHIP）来识别与HUR相关的消息子集并在P19神经元发育过程中研究其动态。 2。研究神经元中HUR和miRNA之间的相互作用。 RIP-CHIP分析以及实时PCR和Northern Analysis将用于识别与神经元中与HUR MRNP相关的miRNA的子集。 3。研究HUR在神经元发展过程中的转录后作用。该目标将研究HUR对神经元发育过程中WNT途径的表达和调节的影响。通过过表达和/或击倒P19细胞中的HUR，并查看对Wnt家族成员RNA以及蛋白质表达和转录活性的影响，我们可以开始理解HUR的调节作用。相关性：该项目将调查神经元发展期间WNT途径的转录后调节。通过更好地理解这种经常被忽视的调节形式，我们将确定涉及WNT途径的疾病的治疗干预措施的潜在靶标。