Intrinsically disordered sequences enable modulation of protein phase separation through distributed tyrosine motifs

Yuan Lin, Simon L. Currie, Michael K. Rosen

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Liquid-liquid phase separation (LLPS) is thought to contribute to the establishment of many biomolecular condensates, eukaryotic cell structures that concentrate diverse macromolecules but lack a bounding membrane. RNA granules control RNA metabolism and comprise a large class of condensates that are enriched in RNA-binding proteins and RNA molecules. Many RNA granule proteins are composed of both modular domains and intrinsically disordered regions (IDRs) having low amino acid sequence complexity. Phase separation of these molecules likely plays an important role in the generation and stability of RNA granules. To understand how folded domains and IDRs can cooperate to modulate LLPS, we generated a series of engineered proteins. These were based on fusions of an IDR derived from theRNAgranule protein FUS (fused in sarcoma) to a multivalent poly-Src homology 3 (SH3) domain protein that phase-separates when mixed with a poly-proline-rich-motif (polyPRM) ligand. We found that the wild-type IDR promotes LLPS of the polySH3-polyPRM system, decreasing the phase separation threshold concentration by 8-fold. Systematic mutation of tyrosine residues in Gly/Ser-Tyr-Gly/Ser motifs of the IDR reduced this effect, depending on the number but not on the position of these substitutions. Mutating all tyrosines to nonaromatic residues or phosphorylating the IDR raised the phase separation threshold above that of the unmodified polySH3- polyPRM pair. These results show that low-complexity IDRs can modulate LLPS both positively and negatively, depending on the degree of aromaticity and phosphorylation status. Our findings provide plausible mechanisms by which these sequences could alter RNA granule properties on evolutionary and cellular timescales.

Original languageEnglish (US)
Pages (from-to)19110-19120
Number of pages11
JournalJournal of Biological Chemistry
Volume292
Issue number46
DOIs
StatePublished - Jan 1 2017

Fingerprint

Phase separation
Tyrosine
Modulation
RNA
Proline
Liquids
Proteins
RNA-Binding Proteins
src Homology Domains
RNA Stability
Eukaryotic Cells
Sarcoma
Amino Acid Sequence
Phosphorylation
Molecules
Ligands
Macromolecules
Mutation
Metabolism
Membranes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Intrinsically disordered sequences enable modulation of protein phase separation through distributed tyrosine motifs. / Lin, Yuan; Currie, Simon L.; Rosen, Michael K.

In: Journal of Biological Chemistry, Vol. 292, No. 46, 01.01.2017, p. 19110-19120.

Research output: Contribution to journalArticle

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