De novo design of RNA-binding proteins with a prion-like domain related to ALS/FTD proteinopathies

Kana Mitsuhashi, Daisuke Ito, Kyoko Mashima, Munenori Oyama, Shinichi Takahashi, Norihiro Suzuki

Research output: Contribution to journalArticle

Abstract

Aberrant RNA-binding proteins form the core of the neurodegeneration cascade in spectrums of disease, such as amyotrophic lateral sclerosis (ALS)/frontotemporal dementia (FTD). Six ALS-related molecules, TDP-43, FUS, TAF15, EWSR1, heterogeneous nuclear (hn)RNPA1 and hnRNPA2 are RNA-binding proteins containing candidate mutations identified in ALS patients and those share several common features, including harboring an aggregation-prone prion-like domain (PrLD) containing a glycine/serine-tyrosine-glycine/serine (G/S-Y-G/S)-motif-enriched low-complexity sequence and rich in glutamine and/or asparagine. Additinally, these six molecules are components of RNA granules involved in RNA quality control and become mislocated from the nucleus to form cytoplasmic inclusion bodies (IBs) in the ALS/FTD-affected brain. To reveal the essential mechanisms involved in ALS/FTD-related cytotoxicity associated with RNA-binding proteins containing PrLDs, we designed artificial RNA-binding proteins harboring G/S-Y-G/S-motif repeats with and without enriched glutamine residues and nuclear-import/export-signal sequences and examined their cytotoxicity in vitro. These proteins recapitulated features of ALS-linked molecules, including insoluble aggregation, formation of cytoplasmic IBs and components of RNA granules, and cytotoxicity instigation. These findings indicated that these artificial RNA-binding proteins mimicked features of ALS-linked molecules and allowed the study of mechanisms associated with gain of toxic functions related to ALS/FTD pathogenesis.

Original languageEnglish
Article number16871
JournalScientific Reports
Volume7
Issue number1
DOIs
Publication statusPublished - 2017 Dec 1

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RNA-Binding Proteins
Prions
Inclusion Bodies
Amyotrophic Lateral Sclerosis
RNA
Glutamine
Glycine
Serine
Nuclear Export Signals
Cell Nucleus Active Transport
Poisons
Asparagine
Protein Sorting Signals
Quality Control
Tyrosine
Frontotemporal Dementia With Motor Neuron Disease
Mutation
Brain
Proteins

ASJC Scopus subject areas

  • General

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De novo design of RNA-binding proteins with a prion-like domain related to ALS/FTD proteinopathies. / Mitsuhashi, Kana; Ito, Daisuke; Mashima, Kyoko; Oyama, Munenori; Takahashi, Shinichi; Suzuki, Norihiro.

In: Scientific Reports, Vol. 7, No. 1, 16871, 01.12.2017.

Research output: Contribution to journalArticle

Mitsuhashi, Kana ; Ito, Daisuke ; Mashima, Kyoko ; Oyama, Munenori ; Takahashi, Shinichi ; Suzuki, Norihiro. / De novo design of RNA-binding proteins with a prion-like domain related to ALS/FTD proteinopathies. In: Scientific Reports. 2017 ; Vol. 7, No. 1.
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