Berlin 2024 – scientific programme
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BP: Fachverband Biologische Physik
BP 27: Single Molecule Biophysics
BP 27.5: Talk
Thursday, March 21, 2024, 10:30–10:45, H 0112
A single-molecule perspective on the RNA interactions of a Ser-Arg-rich splicing factor — •Marie Synakewicz1, Sarah Habeler1, Steffen Winkler1, Lucia Franchini1, Héloïse Bürgisser2, Antoine Cléry1, Frédéric Allain2, Nina Hartrampf1, and Benjamin Schuler1 — 1University of Zurich, Zürich, Switzerland — 2ETH Zurich, Zürich, Switzerland
Boundaries between coding and non-coding regions within mRNAs are recognised by serine-arginine-rich splicing factors (SRSFs) that contain one or two structured RNA recognition motifs (RRMs) and a long intrinsically disordered domain consisting of many Arg-Ser repeats. Extensive phosphorylation of RS domains modulates SRSF conformation, cellular localisation and function. Using single-molecule techniques, we aim to understand the molecular detail of the SRSF1-RNA interaction, and how this is regulated by the RS domain and its phosphorylation pattern. We characterised the interaction of full-length SRSF1, the RRMs and the RS domain with ssRNAs using single-molecule Förster Resonance Energy Transfer (smFRET), before showing that an increase in phosphorylation of the RS domain correlates with a decrease in affinity. More recently, we started to explore protein-RNA interactions in the context of a natural pre-mRNA construct. Using both smFRET and force spectroscopy we show that the conformational ensemble of the pre-mRNA consists of more than one structure, and that these are modulated by protein binding. Our results provide new insights into how SRSF1 can bind and modulate RNA structure, and therefore its capacity to regulate many cellular processes.
Keywords: optical tweezers; single-molecule FRET; RNA binding; RNA structure