We combine solution NMR, X-ray crystallography, small angle scattering, and cryo-EM in employ integrative structural biology to study multidomain RNA-binding proteins (RBPs) and their complexes in RNA-based gene regulation. Specific methyl-labeling and TROSY are combined with PRE and RDC data, and complemented with SAXS experiments. Ultrahigh magnetic fields, i.e. at 1.2 GHz, provide unprecedented resolution, access to conformational dynamics and magnetic field alignment of large RNAs.
Alternative splicing of pre-mRNAs is regulated by multidomain RBPs, which mediate protein-protein and protein-RNA interactions, and further modulated by RNA structure and posttranscriptional modifications. Recent results will be presented that describe how we use NMR to reveal how multidomain RNA binding proteins such as FUBP1, PRP40 and RBM5 mediate protein-RNA and protein-protein interactions for pre-mRNA recognition and bridging of 3’ and 5’ splice sites during spliceosome assembly. We present an unprecedented mode of proline-rich motif recognition by the PRP40 tandem WW domains and regulation of their binding selectivity by intramolecular interactions.