Controlled by disorder: Phosphorylation modulates SRSF1 domain availability for spliceosome assembly.

Abstract

Serine/arginine-rich splicing factor 1 (SRSF1) is key in the mRNA lifecycle including transcription, splicing, nonsense-mediated decay, and nuclear export. Consequently, its dysfunction is linked to cancers, viral evasion, and developmental disorders. The functionality of SRSF1 relies on its interactions with other proteins and RNA molecules. These processes are regulated by phosphorylation of its unstructured arginine/serine-rich tail (RS). Here, we characterize how phosphorylation affects SRSF1's protein and RNA interaction and phase separation. Using NMR paramagnetic relaxation enhancement and chemical shift perturbation, we find that when unphosphorylated, SRSF1's RS interacts with its first RNA-recognition motif (RRM1). Phosphorylation of RS decreases its interactions with the protein-binding site of RRM1 and increases its interactions with the RNA-binding site of RRM1. This change in SRSF1's intramolecular interactions increases the availability of protein-interacting sites on RRM1 and weakens RNA binding of SRSF1. Phosphorylation alters the phase separation of SRSF1 by diminishing the role of arginine in intermolecular interactions. These findings provide an unprecedented view of how SRSF1 influences the early-stage spliceosome assembly.