Phase separation and toxicity of c9orf72 poly(Pr) depends on alternate distribution of arginine

Chen Chen, Yoshiaki Yamanaka, Koji Ueda, Peiying Li, Tamami Miyagi, Yuichiro Harada, Sayaka Tezuka, Satoshi Narumi, Masahiro Sugimoto, Masahiko Kuroda, Yuhei Hayamizu, Kohsuke Kanekura

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19 Citations (Scopus)


Arg (R)-rich dipeptide repeat proteins (DPRs; poly(PR): Pro-Arg and poly(GR): Gly-Arg), encoded by a hexanucleotide expansion in the C9ORF72 gene, induce neurodegeneration in amyotrophic lateral sclerosis (ALS). Although R-rich DPRs undergo liquid–liquid phase separation (LLPS), which affects multiple biological processes, mechanisms underlying LLPS of DPRs remain elusive. Here, using in silico, in vitro, and in cellulo methods, we determined that the distribution of charged Arg residues regulates the complex coacervation with anionic peptides and nucleic acids. Proteomic analyses revealed that alternate Arg distribution in poly(PR) facilitates entrapment of proteins with acidic motifs via LLPS. Transcription, translation, and diffusion of nucleolar nucleophosmin (NPM1) were impaired by poly(PR) with an alternate charge distribution but not by poly(PR) variants with a consecutive charge distribution. We propose that the pathogenicity of R-rich DPRs is mediated by disturbance of proteins through entrapment in the phase-separated droplets via sequence-controlled multivalent protein–protein interactions.

Original languageEnglish
Article numbere202103160
JournalJournal of Cell Biology
Issue number11
Publication statusPublished - 2021 Nov 1
Externally publishedYes

ASJC Scopus subject areas

  • Cell Biology


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