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PLoS One. 2017 Apr 28;12(4):e0176462. doi: 10.1371/journal.pone.0176462. eCollection 2017.

Altered nucleocytoplasmic proteome and transcriptome distributions in an in vitro model of amyotrophic lateral sclerosis.

Author information

1
Department of Neurology, Seoul Medical Center, Seoul, Republic of Korea.
2
Department of Neurology, Seoul National University Seoul Metropolitan Government Boramae Medical Center, Seoul, Republic of Korea.
3
Division of Mass Spectrometry Research, Korea Basic Science Institute, Daejun, Korea.
4
Department of Neurology, Seoul National University Hospital, Seoul, Korea.
5
Macrogen Inc., Seoul, Korea.
6
Department of Neurology, Inha University Hospital, Incheon, Korea.
7
Department of Neurology, Eulji University Hospital, Daejun, Korea.
8
Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul, Korea.

Abstract

Aberrant nucleocytoplasmic localization of proteins has been implicated in many neurodegenerative diseases. Evidence suggests that cytoplasmic mislocalization of nuclear proteins such as transactive response DNA-binding protein 43 (TDP-43) and fused in sarcoma (FUS) may be associated with neurotoxicity in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration. This study investigated the changes in nucleocytoplasmic distributions of the proteome and transcriptome in an in vitro model of ALS. After subcellular fractionation of motor neuron-like cell lines expressing wild-type or G93A mutant hSOD1, quantitative mass spectrometry and next-generation RNA sequencing (RNA-seq) were performed for the nuclear and cytoplasmic compartments. A subset of the results was validated via immunoblotting. A total of 1,925 proteins were identified in either the nuclear or cytoplasmic fractions, and 32% of these proteins were quantified in both fractions. The nucleocytoplasmic distribution of 37 proteins was significantly changed in mutant cells with nuclear and cytoplasmic shifts in 13 and 24 proteins, respectively (p<0.05). The proteins shifted towards the nucleus were enriched regarding pathways of RNA transport and processing (Dhx9, Fmr1, Srsf3, Srsf6, Tra2b), whereas protein folding (Cct5, Cct7, Cct8), aminoacyl-tRNA biosynthesis (Farsb, Nars, Txnrd1), synaptic vesicle cycle (Cltc, Nsf), Wnt signalling (Cltc, Plcb3, Plec, Psmd3, Ruvbl1) and Hippo signalling (Camk2d, Plcb3, Ruvbl1) pathways were over-represented in the proteins shifted to the cytoplasm. A weak correlation between the changes in protein and mRNA levels was found only in the nucleus, where mRNA was relatively abundant in mutant cells. This study provides a comprehensive dataset of the nucleocytoplasmic distribution of the proteome and transcriptome in an in vitro model of ALS. An integrated analysis of the nucleocytoplasmic distribution of the proteome and transcriptome demonstrated multiple candidate pathways including RNA processing/transport and protein synthesis and folding that may be relevant to the pathomechanism of ALS.

PMID:
28453527
PMCID:
PMC5409181
DOI:
10.1371/journal.pone.0176462
[Indexed for MEDLINE]
Free PMC Article

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