Conformational control of multi-domain RNA binding proteins (B03)

多结构域蛋白复合物在早期剪接体组装过程中的结构机制和动力学将被研究。基于我们最近的研究结果，我们将研究3 ‘剪接位点RNA识别过程中的构象开关（涉及必要的剪接因子U2AF2/1， hnRNP A1和SF1），这些因子通过复合物E中的桥接因子与5 ’剪接位点上的U1 snRNP的相互作用，以及参与神经元微外显子识别的蛋白质复合物。综合结构生物学在溶液中，最先进的核磁共振波谱，SAXS/SANS和FRET实验将结合晶体学和冷冻电镜。RNA序列变异、构象平衡、内在无序区域和剪接因子中的疾病相关突变的影响将被表征。这些结果将揭示多结构域蛋白复合物的构象调控和动力学如何影响RNA识别和剪接体组装过程中pre-mRNA剪接的调控。

Structural mechanisms and dynamics of multidomain protein complexes during early spliceosome assembly will be investigated. Building on our recent results, we will study conformational switches during recognition of the 3’ splice site RNA (involving the essential splicing factors U2AF2/1, hnRNP A1 and SF1), the interaction of these factors with U1 snRNP at the 5’ splice site via bridging factors in complex E, and the protein complex involved in recognition of neuronal microexons. Integrative structural biology in solution, state-of-the-art NMR spectroscopy, SAXS/SANS and FRET experiments will be combined with crystallography and cryo-EM. Effects of RNA sequence variations, conformational equilibria, intrinsically disordered regions and disease-linked mutations in the splicing factors will be characterised. The results will unravel how conformational regulation and dynamics of multidomain protein complexes affects RNA recognition and the regulation of pre-mRNA splicing during spliceosome assembly.