Interfering with protein-protein interactions in the spliceosme

干扰剪接体中蛋白质-蛋白质相互作用

• 批准号: 164231578
• 负责人: Professor Dr. Reinhard Lührmann
• 金额: --
• 依托单位: Arbeitsgruppe Strukturbiochemie
• 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 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/164231578?language=en
• 关键词: Interfering; protein; interactions; spliceosme

Pre-mRNA splicing is carried out by an elaborate ribonucleoprotein (RNP) enzyme, the spliceosome, which is assembled anew on each pre-mRNA intron by the stepwise interaction of snRNPs and multiple non-snRNP splicing factors. During its maturation into a catalytically active RNP, the spliceosome is repeatedly remodeled, undergoing numerous compositional and conformational changes. Spliceosome maturation and catalysis are accompanied by profound rearrangements in the spliceosomal RNA and RNP networks and by the dynamic exchanges of more than 100 proteins. At present we have incomplete knowledge of the role of individual proteins or protein complexes and of the sequence of protein exchanges during spliceosome maturation and catalysis. Most likely, a large number of assembly intermediates exist in addition to the limited number of intermediates identified to date. In order to access novel and compositionally homogenous intermediates, we plan to develop and characterize small molecule inhibitors that interfere with particular spliceosomal protein-protein interactions. Inhibitors will be identified by three strategies: (1) Rational design based on crystal structures of protein complexes; (2) Targeted screening based on isolated protein pairs; (3) Targeted screening based on a protein recruitment assay. Initial hits will be improved by synthetic optimization aided by co-crystal structures of proteins in complex with inhibitors. The results obtained will not only help to elucidate the conformational dynamics of spliceosome assembly and catalysis but may also provide useful leads for the development of therapeutic spliceosome inhibitors.

前体mRNA剪接是由一种复杂的核糖核蛋白（RNP）酶剪接体进行的，剪接体通过snRNP和多种非snRNP剪接因子的逐步相互作用在每个前体mRNA内含子上重新组装。在其成熟为具有催化活性的RNP期间，剪接体反复重塑，经历许多组成和构象变化。剪接体成熟和催化伴随着剪接体RNA和RNP网络中的深刻重排以及100多种蛋白质的动态交换。目前，我们对单个蛋白质或蛋白质复合物的作用以及剪接体成熟和催化过程中蛋白质交换的顺序还不完全了解。最有可能的是，除了迄今为止确定的有限数量的中间体之外，还存在大量的组装中间体。为了获得新的和组成上均匀的中间体，我们计划开发和表征干扰特定剪接体蛋白质-蛋白质相互作用的小分子抑制剂。将通过三种策略鉴定抑制剂：（1）基于蛋白质复合物的晶体结构的合理设计;（2）基于分离的蛋白质对的靶向筛选;（3）基于蛋白质募集测定的靶向筛选。通过与抑制剂复合的蛋白质的共晶体结构辅助的合成优化，将改善初始命中。所获得的结果不仅有助于阐明剪接体组装和催化的构象动力学，而且还可能为开发治疗性剪接体抑制剂提供有用的线索。

项目成果

Structural basis for dual roles of Aar2p in U5 snRNP assembly.

Aar2p 在 U5 snRNP 组装中双重作用的结构基础

• DOI: 10.1101/gad.213207.113
• 发表时间: 2013
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: Cristao;Santos;Mozaffari Jovin;Heroven;Holton;Lührmann;Wahl M.C.
• 通讯作者: Wahl M.C.

Molecular architecture of zinc chelating small molecules that inhibit spliceosome assembly at an early stage.

早期抑制剪接体组装的锌螯合小分子的分子结构

• DOI: 10.1261/rna.034819.112
• 发表时间: 2012
• 期刊: RNA
• 影响因子: 4.5
• 作者: Canzoneri;Samatov;Lührmann;Oyelere
• 通讯作者: Oyelere

Mechanism for Aar2p function as a U5 snRNP assembly factor.

• DOI: 10.1101/gad.635911
• 发表时间: 2011-08
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: G. Weber;Vanessa F. Cristão;Flavia de L Alves;K. Santos;N. Holton;J. Rappsilber;J. Beggs;M. Wahl