Mutant human FUS Is ubiquitously mislocalized and generates persistent stress granules in primary cultured transgenic zebrafish cells

PLoS One. 2014 Jun 9;9(6):e90572. doi: 10.1371/journal.pone.0090572. eCollection 2014.

Abstract

FUS mutations can occur in familial amyotrophic lateral sclerosis (fALS), a neurodegenerative disease with cytoplasmic FUS inclusion bodies in motor neurons. To investigate FUS pathology, we generated transgenic zebrafish expressing GFP-tagged wild-type or fALS (R521C) human FUS. Cell cultures were made from these zebrafish and the subcellular localization of human FUS and the generation of stress granule (SG) inclusions examined in different cell types, including differentiated motor neurons. We demonstrate that mutant FUS is mislocalized from the nucleus to the cytosol to a similar extent in motor neurons and all other cell types. Both wild-type and R521C FUS localized to SGs in zebrafish cells, demonstrating an intrinsic ability of human FUS to accumulate in SGs irrespective of the presence of disease-associated mutations or specific cell type. However, elevation in relative cytosolic to nuclear FUS by the R521C mutation led to a significant increase in SG assembly and persistence within a sub population of vulnerable cells, although these cells were not selectively motor neurons.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Cells, Cultured
  • Cytoplasmic Granules / metabolism*
  • Cytosol / metabolism*
  • Humans
  • Motor Neurons / cytology
  • Mutation*
  • Protein Transport / genetics
  • RNA-Binding Protein FUS / genetics*
  • RNA-Binding Protein FUS / metabolism*
  • Zebrafish / genetics*
  • Zebrafish / metabolism*

Substances

  • RNA-Binding Protein FUS

Grant support

Funding provided by The National Health and Medical Research Council (NHMRC), Australia, The Rebecca Cooper Medical Research Foundation and The MND Research Institute of Australia The Snow Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.