TY - JOUR
T1 - Phase separation and toxicity of c9orf72 poly(Pr) depends on alternate distribution of arginine
AU - Chen, Chen
AU - Yamanaka, Yoshiaki
AU - Ueda, Koji
AU - Li, Peiying
AU - Miyagi, Tamami
AU - Harada, Yuichiro
AU - Tezuka, Sayaka
AU - Narumi, Satoshi
AU - Sugimoto, Masahiro
AU - Kuroda, Masahiko
AU - Hayamizu, Yuhei
AU - Kanekura, Kohsuke
N1 - Funding Information:
This work was supported by grants from the Japan Society for the Promotion of Science KAKENHI (16H06247, 17H03923, and 20H03593 to K. Kanekura; 17K15671 to Y. Harada; 16H05973 and 20H02564 to Y. Hayamizu; and 17H04067 to M. Kuroda). This work was also supported in part by the Japan Agency for Medical Research and Development (16ek0109180h0001 and 17ae0101016s0904), Strategic Research Foundation Grant-aided Project for Private Universities from the Ministry of Education, Culture, Sports, Science and Technology of Japan (M. Kuroda), Takeda Science Foundation (K. Kanekura), Japan Intractable Diseases (Nanbyo) Research Foundation (K. Kanekura), the Tokyo Biochemical Research Foundation (K. Kanekura), and the Ichiro Kanehara Foundation (K. Kanekura). The authors declare no competing financial interests.
Publisher Copyright:
© 2021 Chen et al.
PY - 2021/11/1
Y1 - 2021/11/1
N2 - 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.
AB - 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.
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U2 - 10.1083/jcb.202103160
DO - 10.1083/jcb.202103160
M3 - Article
C2 - 34499080
AN - SCOPUS:85115958155
SN - 0021-9525
VL - 220
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 11
M1 - e202103160
ER -