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Mammalian Expression> | … All peptides were chemically synthesized by Genscript with purity >85%. Human rRNA was extracted from HEK293 cells using the RNeasy kit (Qiagen). Human NPM1 cDNA was amplified from NPM1-GFP (gift of Xin Wang, Center for Cancer Research, National Cancer Institute, … | Get A Quote |
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, ... More