NMR and integrative structural biology to study dynamic molecular interactions – from molecular chaperones to splicing regulation

Michael Sattler

We combine solution NMR, X-ray crystallography, small angle scattering, and cryo-EM in integrative structural biology to study multidomain (RNA-binding) proteins and protein complexes that play important roles in RNA-based gene regulation, cellular signalling and molecular chaperones. NMR is an important, versatile method to study molecular interactions and conformational dynamics of protein, RNAs and their complexes and for structure determination of small and dynamics complexes, which are difficult to study by crystallography and cryo-EM. NMR methods can be readily employed for validation of structural models predicted by deep learning methods, such as AlphaFold2.

The lecture will highlight NMR approaches available for such studies, providing examples from recent applications in our lab that highlight dynamic protein-RNA interactions, RNA structure and dynamic conformational states in molecular chaperones and benefits of ultrahigh-magnetic fields at 1.2 GHz proton Larmor frequency.

References:

• Ebersberger S, Hipp C, Mulorz MM, et al Luck K, Sattler M, Konig J
  FUBP1 is a general splicing factor facilitating 3' splice site recognition and splicing of long introns.
  (2023) Mol Cell 83, 2653-2672 e2615. doi:10.1016/j.molcel.2023.07.002

• Soni K, Jagtap PKA, Martinez-Lumbreras S, Bonnal S, Geerlof A, Stehle R, Simon B, Valcarcel J, Sattler M
  Structural basis for specific RNA recognition by the alternative splicing factor RBM5.
  (2023) Nat Commun 14, 4233. doi:10.1038/s41467-023-39961-w

• Jones AN, Grass C, Meininger I, Geerlof A, Klostermann M, Zarnack K, Krappmann D, Sattler M
  Modulation of pre-mRNA structure by hnRNP proteins regulates alternative splicing of MALT1.
  (2022) Sci Adv 8, eabp9153. doi:10.1126/sciadv.abp9153

• Biebl MM, Delhommel F, Faust O, Zak KM, Agam G, Guo X, Muhlhofer M, Dahiya V, Hillebrand D, Popowicz GM, Kampmann M, Lamb DC, Rosenzweig R, Sattler M, Buchner J
  NudC guides client transfer between the Hsp40/70 and Hsp90 chaperone systems.
  (2022) Mol Cell 82, 555-569 e7. doi:10.1016/j.molcel.2021.12.031

• Lopez A, Dahiya V, Delhommel F, Freiburger L, Stehle R, Asami S, Rutz D, Blair L, Buchner J, Sattler M
  Client binding shifts the populations of dynamic Hsp90 conformations through an allosteric network.
  (2021) Sci Adv 7. doi:10.1126/sciadv.abl7295