Mechanism of action of an lncRNA for directing muscle differentiation

lncRNA指导肌肉分化的作用机制

基本信息

批准号：
10396900
负责人：
Anindya Dutta
金额：
$ 21.39万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10396900
关键词：
Mechanism action lncRNA directing muscle

项目摘要

DESCRIPTION (provided by applicant): Long noncoding RNAs (lncRNAs) are noncoding long (>100 bases) single- stranded RNAs that have burst onto the scene as major regulators of gene expression. MicroRNAs, are about 22 base long single-stranded RNAs that came to the fore about a decade back and this laboratory has been among the pioneers that demonstrated the roles of several microRNAs in the differentiation of skeletal muscle myoblasts to myotubes. We now propose to use our expertise in the role of RNAs in muscle differentiation to determine the mechanism by which one particular muscle- differentiation-induced (MDI) lncRNA, MUNC, regulates gene expression and promotes muscle differentiation. Aim 1 will test the hypothesis that MUNC is essential for muscle differentiation because of its role in either regulating the binding or the expression of critical myogenic transcription factors as a constituent of novel protein complexes involved in gene regulation. Aim 2 will carry out structure function studies on mouse and human MUNC to test the hypothesis that evolutionarily conserved domains exist in this lncRNA that are critical for its mechanism of action. Aim 3 will identify new protein and DNA partners of MUNC to identify functions of MUNC independent of Myogenic transcription factors. This Aim will also identify additional lncRNAs that are induced during muscle differentiation and integrate their role in differentiation with that of MUNC and MyoD. This is an exciting project that involves the collaboration of a molecular biologist with a skeletal muscle physiologist. It wil use new experimental reagents such as lncRNAs and state-of-the-art approaches like ChIRP. LC-MS-MS, SILAC and ChiRP-seq to discover the mechanism of action of one lncRNA in control of cell-type specificity. Since our preliminary results identify tens of novel lncRNAs induced during differentiation, at least some of which already implicated in differentiation, success of this project will launch a prolonged investigation of several lncRNAs in this very important paradigm of differentiation.

描述（由适用提供）：长的非编码RNA（LNCRNA）是非编码的长（> 100个碱基）的单链RNA，它们已成为基因表达的主要调节剂。 MicroRNA是大约22个长的单链RNA，大约十年后出现，并且该实验室一直是一些先驱者之一，这些先驱证明了几种microRNA在骨骼肌成肌细胞与肌管中的分化中的作用。现在，我们建议在RNA在肌肉分化中的作用中使用我们的专业知识来确定一种特定的肌肉分化诱导的（MDI）lncRNA的机制，MUNC调节基因表达并促进肌肉分化。 AIM 1将检验以下假设：MUNC对于肌肉分化至关重要，因为它在衡量关键的肌源性转录因子的结合或表达中的作用是构成与基因调节有关的新型蛋白质复合物的构成的。 AIM 2将对小鼠和人类MUNC进行结构函数研究，以检验以下假设：在该lncRNA中存在进化构成的结构域对于其作用机理至关重要。 AIM 3将确定MUNC的新蛋白质和DNA伴侣，以鉴定MUNC的功能，而不是肌源性转录因子。该目标还将确定在肌肉分化过程中诱导的其他LNCRNA，并将其与Munc和Myod的分化相结合。这是一个令人兴奋的项目，涉及分子生物学家与骨骼肌肉生理学家的合作。它将使用新的实验试剂，例如LNCRNA和CHIRP等最先进的方法。 LC-MS-MS，SILAC和CHIRP-SEQ发现一个LNCRNA控制细胞类型特异性的作用机理。由于我们的初步结果确定了在分化过程中诱导的数十种新型LNCRNA，至少有一些已经与分化有关，因此该项目的成功将在这种非常重要的分化范式中延长对几个LNCRNA的研究。