Open AccessPhase separation and toxicity of C9orf72 poly(PR) depends on alternate distribution of arginine
Author(s) -
Chen Chen,
Yoshiaki Yamanaka,
Koji Ueda,
Peiying Li,
Tamami Miyagi,
Yuichiro Harada,
Sayaka Tezuka,
Shunji Narumi,
Masahiro Sugimoto,
Masahiko Kuroda,
Yuhei Hayamizu,
Kohsuke Kanekura
Publication year - 2021
Publication title -
the journal of cell biology/the journal of cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.414
H-Index - 380
eISSN - 1540-8140
pISSN - 0021-9525
DOI - 10.1083/jcb.202103160
Subject(s) - biology , nucleophosmin , intrinsically disordered proteins , microbiology and biotechnology , c9orf72 , biophysics , neurodegeneration , biochemistry , trinucleotide repeat expansion , gene , medicine , allele , disease , pathology
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.