Non-coding RNAs in Transcription and Chromatin Architecture

描述（由申请人提供）： 我们最近发现了一类被称为ncRNA-activating（ncRNA-a）的长ncRNA，它可以在长距离上激活邻近蛋白质编码基因的表达。我们目前的数据表明，ncRNA-a通过与辅激活因子复合物Mediator结合来促进DNA循环。将研究ncRNA-a和介体复合物之间的关联，以了解如何实现靶选择的特异性，并查明RNA对转录激活的贡献。 这三个目标旨在提供对激活ncRNA及其与介体复合物的关联的全面分子理解。Aim 1描述了研究ncRNA-a核定位和转录激活的分子基础的实验。目的2描述了描述ncRNA-a和介体复合物相互作用的分子基础的实验。我们将确定ncRNA-a是否对介体募集到其靶位点至关重要，并进行结构/功能实验以评估ncRNA-a/介体相互作用在转录激活中的重要性。Aim 3将评估ncRNA-a激活CDK 8激酶活性的分子基础。重要的是，介体复合物Med 12亚基的复发性突变，这是激酶活性的关键，导致多种人类认知综合征和良性平滑肌肿瘤的发展。因此，我们将检验这样的假设，即这些突变通过破坏其关联来影响Mediator的激酶活性。 与ncRNA-a。

DESCRIPTION (provided by applicant): Project Summary We have recently identified a class of long ncRNA termed ncRNA-activating (ncRNA-a), which can activate the expression of neighboring protein-coding genes at long distances. Our current data suggest that ncRNA-a function by associating with the co-activator complex, Mediator, to promote DNA looping. The association between ncRNA-a and the Mediator complex will be investigated to understand how specificity for target selection is achieved and to pinpoint the contribution of RNA to transcriptional activation. The three Aims are designed to provide a thorough molecular understanding of the activating ncRNAs and their association with the Mediator complex. Aim1 describes experiments that examine the molecular basis for ncRNA-a nuclear localization and transcriptional activation. Aim 2 describes experiments to delineate the molecular basis of ncRNA-a and the Mediator complex interaction. We will determine whether ncRNA-a is critical for the recruitment of Mediator to its target sites and perform structure/function experiments to assess the importance of ncRNA-a/Mediator interaction in transcriptional activation. Aim3 will assess the molecular basis by which ncRNA-a activates the CDK8 kinase activity. Importantly, recurrent mutations in Med12 subunit of Mediator complex, which is critical for kinase activity, result in development of multiple human cognitive syndromes and a benign smooth muscle neoplasm. Therefore, we will test the hypothesis that such mutations affect the kinase activity of Mediator by disrupting its association with the ncRNA-a.