Peptides that Mimic RS repeats modulate phase separation of SRSF1, revealing a reliance on combined stacking and electrostatic interactions

Elife. 2023 Mar 2:12:e84412. doi: 10.7554/eLife.84412.


Phase separation plays crucial roles in both sustaining cellular function and perpetuating disease states. Despite extensive studies, our understanding of this process is hindered by low solubility of phase-separating proteins. One example of this is found in SR and SR-related proteins. These proteins are characterized by domains rich in arginine and serine (RS domains), which are essential to alternative splicing and in vivo phase separation. However, they are also responsible for a low solubility that has made these proteins difficult to study for decades. Here, we solubilize the founding member of the SR family, SRSF1, by introducing a peptide mimicking RS repeats as a co-solute. We find that this RS-mimic peptide forms interactions similar to those of the protein's RS domain. Both interact with a combination of surface-exposed aromatic residues and acidic residues on SRSF1's RNA Recognition Motifs (RRMs) through electrostatic and cation-pi interactions. Analysis of RRM domains from human SR proteins indicates that these sites are conserved across the protein family. In addition to opening an avenue to previously unavailable proteins, our work provides insight into how SR proteins phase separate and participate in nuclear speckles.

Keywords: NMR; SR proteins; SRSF1; cation-pi interaction; human; intrinsically disordered protein; molecular biophysics; phase separation; structural biology.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, N.I.H., Extramural

MeSH terms

  • Alternative Splicing*
  • Arginine / metabolism
  • Humans
  • Nuclear Proteins* / metabolism
  • Peptides / metabolism
  • Phosphorylation
  • RNA Splicing
  • Serine / metabolism
  • Serine-Arginine Splicing Factors / genetics
  • Serine-Arginine Splicing Factors / metabolism
  • Static Electricity


  • Nuclear Proteins
  • Peptides
  • Arginine
  • Serine
  • SRSF1 protein, human
  • Serine-Arginine Splicing Factors