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Mol Biol Cell. 2014 Sep 1;25(17):2571-8. doi: 10.1091/mbc.E14-05-1007. Epub 2014 Jul 9.

FUS is sequestered in nuclear aggregates in ALS patient fibroblasts.

Author information

1
Howard Hughes Medical Institute, Department of Chemistry and Biochemistry and BioFrontiers Institute, University of Colorado, Boulder, CO 80309.
2
Department of Neurology, Massachusetts General Hospital, Boston, MA 02114 Howard Hughes Medical Institute, Harvard Stem Cell Institute and Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138.
3
Department of Neurology, Massachusetts General Hospital, Boston, MA 02114.
4
Howard Hughes Medical Institute, Harvard Stem Cell Institute and Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138.
5
Howard Hughes Medical Institute, Department of Chemistry and Biochemistry and BioFrontiers Institute, University of Colorado, Boulder, CO 80309 thomas.cech@colorado.edu.

Abstract

Mutations in the RNA-binding protein FUS have been shown to cause the neurodegenerative disease amyotrophic lateral sclerosis (ALS). We investigate whether mutant FUS protein in ALS patient-derived fibroblasts affects normal FUS functions in the nucleus. We investigated fibroblasts from two ALS patients possessing different FUS mutations and a normal control. Fibroblasts from these patients have their nuclear FUS protein trapped in SDS-resistant aggregates. Genome-wide analysis reveals an inappropriate accumulation of Ser-2 phosphorylation on RNA polymerase II (RNA Pol II) near the transcription start sites of 625 genes for ALS patient cells and after small interfering RNA (siRNA) knockdown of FUS in normal fibroblasts. Furthermore, both the presence of mutant FUS protein and siRNA knockdown of wild-type FUS correlate with altered distribution of RNA Pol II within fibroblast nuclei. A loss of FUS function in orchestrating Ser-2 phosphorylation of the CTD of RNA Pol II is detectable in ALS patient-derived fibroblasts expressing mutant FUS protein, even when the FUS protein remains largely nuclear. A likely explanation for this loss of function is the aggregation of FUS protein in nuclei. Thus our results suggest a specific mechanism by which mutant FUS can have biological consequences other than by the formation of cytoplasmic aggregates.

PMID:
25009283
PMCID:
PMC4148247
DOI:
10.1091/mbc.E14-05-1007
[Indexed for MEDLINE]
Free PMC Article

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