Analysis of cytoplasmic functions of the exon junction complex

外显子连接复合物的细胞质功能分析

• 批准号: 186127969
• 负责人: Professor Dr. Niels H. Gehring
• 金额: --
• 依托单位: Institut für Genetik
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2010
• 资助国家: 德国
• 起止时间: 2009-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/186127969?language=en
• 关键词: Analysis; cytoplasmic; functions; exon; junction

Splicing of intron containing transcripts in the nucleus of metazoan cells leads to the deposition of exon junction complexes (EJC) on the nascent mRNAs. Within the cytoplasm, EJCs are essential marks for nonsense-mediated mRNA decay (NMD) and increase the translation efficiency of spliced mRNAs. Both effects are likely interconnected by shared components of the EJC and the translational machinery. We have previously identified a critical role of the EJC component BTZ during NMD and want to further elucidate its mechanism of NMD activation. We will characterize target mRNAs regulated by BTZ and identify interacting co-factors that regulate BTZ’s NMD function. The translation stimulation function of the EJC is mediated by the EJC interacting factor SKAR. The mTOR-dependent kinase S6K1 is recruited by a direct protein-protein interaction to EJC-bound SKAR, phosphorylates important components of the mRNP and thereby stimulates the translation efficiency of the bound mRNA. We aim to characterize the molecular mechanism and the EJC components involved in this process and to identify central phosphorylated mRNP effector proteins. Together, these complementary projects will address the mechanism of two important gene expression processes that are molecularly linked at the level of the EJC.

含有转录物中内含子的剪接在后生细胞核中导致外显子连接复合物（EJC）在新生的mRNA上的沉积。在细胞质中，EJC是废话介导的mRNA衰变（NMD）的必要标记，并提高了剪接mRNA的翻译效率。这两种效果都可能与EJC和翻译机械的共享组件相互联系。我们先前已经确定了NMD期间EJC组件BTZ的关键作用，并希望进一步阐明其NMD激活机理。我们将表征由BTZ调节的目标mRNA，并确定调节BTZ NMD函数的相互作用的共同因素。 EJC的翻译刺激功能是由EJC相互作用因子SKAR介导的。 MTOR依赖性激酶S6K1是通过直接蛋白 - 蛋白质相互作用与EJC结合的Skar募集的，磷酸化MRNP的重要成分，从而刺激了结合mRNA的翻译效率。我们旨在表征该过程中涉及的分子机制和EJC成分，并鉴定中央磷酸化的MRNP效应蛋白。总之，这些完整的项目将解决两个重要的基因表达过程的机制，它们在EJC水平上分子链接。

项目成果

CWC22 connects pre-mRNA splicing and exon junction complex assembly.

• DOI: 10.1016/j.celrep.2012.08.017
• 发表时间: 2012-09
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Anna-Lena Steckelberg;Volker Boehm;A. Gromadzka;Niels H. Gehring

Studying the composition of mRNPs in vitro using splicing-competent cell extracts.

使用剪接活性细胞提取物研究体外 mRNP 的组成

• DOI: 10.1016/j.ymeth.2013.08.033
• 发表时间: 2014
• 期刊: Methods
• 影响因子: 4.8
• 作者: Steckelberg AL;Gehring NH
• 通讯作者: Gehring NH